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Advanced Diagnostic
           Technologies in Public
                  Health


                  Dr Abhay Dhanorkar



2/7/2013                               1
Scope
ā€¢ Introduction
ā€¢ IPHS standards for availability of laboratory services at various
  hospitals.
ā€¢ Classification of diagnostics tests
ā€¢ Immunopathology
ā€¢ Molecular Biology
ā€¢ Clinical Chemistry
ā€¢ Haematology
ā€¢ Cytopathology
ā€¢ Histopathology
ā€¢ Radiology
ā€¢ Electrophysiology
ā€¢ Endoscopy
ā€¢ Summary

2/7/2013                                                          2
Introduction

ā€¢ A diagnostic test is any kind of medical test
  performed to aid in the diagnosis or detection of
  disease.
ā€¢ It may be used to diagnose diseases or measure
  the progress or recovery from disease or confirm
  that a person is free from disease.
ā€¢ Some medical tests are parts of a simple physical
  examination which require only simple tools in
  the hands of a skilled practitioner and can be
  performed in an office environment.
2/7/2013                                          3
Introduction contdā€¦ā€¦

 ā€¢ Some other tests require elaborate equipment
   used by medical technologists or the use of a
   sterile operating theatre environment.
 ā€¢ Some tests require samples of tissue or body
   fluids to be sent off to a pathology lab for
   further analysis.
 ā€¢ Some simple chemical tests such as urine pH,
   can be measured directly in the physicians
   office.

 2/7/2013                                      4
Introduction contdā€¦ā€¦


  ā€¢ However, with advancements in health care, a
    large array of diagnostic tests have become
    available. Various modern (and often,
    expensive) diagnostic modalities are often
    routinely used in medical world.
  ā€¢ According to IPHS standards stated as a part of
    NRHM, the some laboratory facilities should
    be available at various levels.

  2/7/2013                                        5
IPHS standards for availability of laboratory
        services at 31 to 50 bedded hospitals




2/7/2013                                          6
For 51-100 Bedded (All as in 31-50 bedded
with the following additional facilities)




2/7/2013                                7
For 101-200 Bedded (All as in 51-100 bedded
 with the following additional facilities)

   Speciality                Diagnostic Services / Tests

                Colonoscopy, Bronchoscopy, Arthroscopy, Laparoscopy
  Endoscopy
                       (Diagnostic), Colposcopy, Hysteroscopy

    Serology    RA factor test CPK, Lepto spirosis (Rapid test / ELISA)

                HSG, Ultrasonography Colour Doppler, Spiral CT scan,
   Radiology
                                 MRI 0.5 TESSLA

 Biochemistry       Glycosylated Hemoglobin, Blood gas analysis
2/7/2013                                                                  8
For 201-300 Bedded (All as in 101-200 bedded
 with the following additional facilities)




 2/7/2013                                  9
For >300 Bedded (All as in 201-300 bedded
  with the following additional facilities)

            Speciality                      Diagnostic Services

                                      Angiography, Multislice CT, Multislice
             Radiology
                                                     MRI

                                       Bone Marrow Biopsy,Pleural Biopsy,
               Biopsy
                                              Pericardial tapping
                                       Gastroscopy (Oesophagus, stomach,
  Endoscopic Specialised Procedures   deudenum), Sigmoid, Laproscopy and
           and Diagnostic               Colonoscopy, Bronchoscopy and
                                             Foreign Body Removal
2/7/2013                                                                   10
Classification

ā€¢ The various modern diagnostic technologies can
  be classified as
      ā€“    Immunopathology
      ā€“    Molecular Biology
      ā€“    Clinical Chemistry
      ā€“    Haematology
      ā€“    Cytopathology
      ā€“    Histopathology
      ā€“    Radiology
      ā€“    Electrophysiology
      ā€“    Endoscopy

2/7/2013                                       11
Diagnostic Technologies in
               Immunopathology
ā€¢ Antibodies are produced in response to
  antigenic stimulation.
ā€¢ Immunoassays (antigen antibody reactions)
  can be used for detection of either antigens or
  antibodies. For detecting antigens, the
  corresponding specific antibody should be
  used as one of the reagents and vice versa.


2/7/2013                                        12
Immunopathology contdā€¦

 The technologies available are ā€“
 ā€¢ Precipitation immunoassays : It is based on the occurrence of
   precipitation when large complexes of antigens and antibodies
   combine to form an insoluble lattice.
   Ex. :- Widal test and the Weil-Felix test.
 ā€¢ Particle immunoassay : Here specific antigens are coated onto a
   particle and on reaction with antibody under test, the
   agglutination is made more visible. Reverse agglutination where
   the antibody is coated onto the particle is also performed. The
   particle used may be RBC, latex or gelatin.
   Ex. :- 1) TPHA (Treponema pallidum Hemagglutination).
             2) Reverse passive hemagglutination (RPHA) for hepatitis B surface antigen (HBsAg).
             3) Latex agglutination is used in measurement of hCG for qualitative pregnancy tests

  2/7/2013                                                                                          13
Immunopathology contdā€¦


  Radioimmunoassay :
  ā€¢ Radioimmunoassay (RIA) is used to test antigens
    (e.g.hormone levels in the blood).
  ā€¢ It is extremely sensitive and extremely specific, but requires a
    sophisticated apparatus and is costly. It also requires special
    precautions, since radioactive substances are used.
  ā€¢ Therefore, today it has been largely supplanted by the ELISA
    method, where the antigen-antibody reaction is measured
    using colorometric signals instead of a radioactive signal.
  ā€¢ To perform a radioimmunoassay, a known quantity of an
    antigen is made radioactive, frequently by labeling it with
    gamma-radioactive isotopes of iodine attached to thyrosine.
  2/7/2013                                                        14
Immunopathology contdā€¦


  ā€¢ This radiolabeled antigen is then mixed with a known
    amount of antibody for that antigen and as a result,
    the two chemically bind to one another.
  ā€¢ Then, a sample of serum from a patient containing an
    unknown quantity of that same antigen is added. This
    causes the unlabeled (or ā€œcoldā€) antigen from the
    serum to compete with the radiolabeled antigen for
    antibody binding sites.
  Applications in public health :
    Ex. Thyroid hormone levels in goiter endemic areas.


  2/7/2013                                            15
Immunopathology contdā€¦


  Enzyme Linked Immunosorbent Assay (ELISA) :
  ā€¢ This is a technique used mainly in immunology to
    detect the presence of an antibody or an antigen
    in a sample.
  ā€¢ In ELISA an unknown amount of antigen is affixed
    to a surface and then a specific antibody is
    washed over the surface so that it can bind to the
    antigen.
  ā€¢ This antibody is linked to an enzyme and in the
    final step a substance is added that the enzyme
    can convert to some detectable signal.
  2/7/2013                                          16
Immunopathology contdā€¦


  Applications in public health :
        ā€“    HIV test or West Nile Virus.
             ELISA can be used for detecting tumour
             markers for certain cancers, e.g. Prostate
             Specific Antigen (PSA) in carcinoma
             prostate.



  2/7/2013                                            17
Immunopathology contdā€¦


  Fluorescent immunoassay :
  ā€¢ When fluorescent molecules are irradiated with light at
     appropriate wavelengths, an electron in the ground state is
     transited into the excited state. As the electron returns to
     the ground state, physical energy is released in the form of
     a photon which is detectable.
  Applications in public health :
  ā€¢ For detecting antigens in tissue sections in kidney biopsies.
  ā€¢ Use of Auramine rhodamine stain in detection of tubercle
     bacilli in smears shown to have better sensitivity than the
     conventional ZN stain.
  ā€¢ To detect the concentration of drugs, hormones and
     proteins and polypeptides.

  2/7/2013                                                     18
Diagnostic Technologies in
              Molecular Biology
Diagnostic Technologies in Molecular Biology
ā€¢ Molecular biology is the study of biology at a molecular
  level.
ā€¢ Molecular biology chiefly concerns itself with
  understanding the interactions between the various
  systems of a cell, including the interactions between
  DNA, RNA and protein biosynthesis and learning how
  these interactions are regulated.
ā€¢ The tools of molecular biology is extremely useful in
  diagnosis, therapy, epidemiologic investigations and
  infection control.

2/7/2013                                                19
Molecular Biology contdā€¦


  ā€¢ Ease of performance, reproducibility, sensitivity
    and specificity of molecular tests are important,
    cost and potential contribution to patient care
    are also of concern.
  ā€¢ Molecular methods may be an improvement over
    conventional microbiologic testing as they are
    most practical and useful application is in
    detecting and identifying infectious agents for
    which routine growth-based culture and
    microscopy methods may not be adequate.

  2/7/2013                                          20
Molecular Biology contdā€¦


  Gel electrophoresis :
  ā€¢ It is one of the principal tools of molecular biology. The
    basic principle is that DNA, RNA and proteins can all be
    separated by means of an electric field.
  ā€¢ In agarose gel electrophoresis, DNA and RNA can be
    separated on the basis of size by running the DNA
    through an agarose gel.
  ā€¢ Proteins can be separated on the basis of size by using
    an SDS-PAGE gel, or on the basis of size and their
    electric charge by using 2D gel electrophoresis.


  2/7/2013                                                  21
Molecular Biology contdā€¦


  Hybridisation assays :
  ā€¢ When the hybridization reaction is used to analyse the
    nucleic acid content of an unknown sample, the
    process is known as a hybridization assay.
  ā€¢ The property of complimentary base pairing allows
    fragments of known composition (the probes) to
    interrogate an unknown for the presence of matching
    (complimentary) sequences.
  ā€¢ The detection of the hybrids can be done a variety of
    technologies     including      radioisotope    labels,
    fluorochrome based detection and enzyme based
    systems.

  2/7/2013                                               22
Molecular Biology contdā€¦


 ā€¢ Liquid phase hybridization : When both sample
   and probe are in solution.
 ā€¢ Solid support hybridization : In these assays,
   hybridization occurs in a biphasic environment, a
   solid phase (usually sample) and a liquid phase
   (usually probe).
 ā€¢ Southern and Northern hybridization assays :
   These combine electrophoretic separation of test
   nucleic acids with transfer to a solid support and
   subsequent hybridization.
 2/7/2013                                          23
Molecular Biology contdā€¦



  ā€¢ The original procedure was described by EM
    Southern and the test nucleic acid was DNA.
    When RNA is the nucleic acid under test, the
    technique is called northern blotting by
    analogy.
  ā€¢ In situ hybridization : This is the detection of
    specific genetic information within a
    morphological context (intact tissue, cells).

  2/7/2013                                         24
Molecular Biology contdā€¦


 Amplification technology :
 ā€¢ Amplification systems are enzyme based processes in
   which a single enzyme or multiple enzymes synthesize
   copies of target nucleic acid.
 ā€¢ All result in production of billions of copies of the
   amplified product in a few hours.
 ā€¢ These techniques are subject to contamination from
   product molecules of previous amplifications and
   hence false positivity is high.
 ā€¢ However, special lab design, practices and workflow
   have helped to reduce false positives to an acceptable
   range.

 2/7/2013                                              25
Molecular Biology contdā€¦
  Polymerase Chain Reaction (PCR)

  ā€¢ It is a simple in vitro chemical reaction that permits
    synthesis of large quantities of nucleic acid.
  ā€¢ This is brought about by heating the reaction mixture
    to separate the strands of the DNA, then cooled to
    permit the primers to anneal to the target DNA in a
    sequence specific manner. The DNA polymerase then
    initiates extension from end.
  ā€¢ Thus the whole process is carried out in a
    programmable thermocycler which controls the
    temperatures at which various steps occur.


  2/7/2013                                              26
Molecular Biology contdā€¦


  ā€¢ Reverse transcriptase PCR (RT-PCR) : PCR was initially
    described to amplify DNA. RT-PCR was developed to
    amplify RNA targets
  ā€¢ Nested PCR : This variation increases the sensitivity
    and specificity of PCR. The products of the first round
    of amplification are subject to second round of
    amplification with a second set of primers.
  ā€¢ Real time PCR : Here the target amplification and
    detection occur simultaneously.
  ā€¢ The computer software supporting the thermocycler
    monitors the data throughout the PCR at every cycle
    and generates a plot.

  2/7/2013                                               27
Molecular Biology contdā€¦



  Applications of molecular biology in Public
    Health
  ā€¢ Pathogen detection by hybridisation : The
    DNA of the infectious agent is detected
    directly in the clinical specimens by DNA
    probes.



  2/7/2013                                  28
Molecular Biology contdā€¦


  ā€¢ DNA probes for culture identification : Probes are available
    for various organisms like Mycobacterium tuberculosis,
    Chlamydia trachomatis, Neisseria gonorrhoeae, Human
    papilloma virus, Hepatitis C virus, Cytomegalovirus and
    Herpes simplex virus.
  ā€¢ This is especially important for slow growing organisms like
    Mycobacteria, fungi etc. Identification of cultured
    Mycobacteria by conventional methods is slow and time
    consuming.
  ā€¢ The use of probes for this purpose permits identification
    from cultures within one working day and sensitivity
    (95.4%) and specificity (99%) are excellent.


  2/7/2013                                                    29
Molecular Biology contdā€¦


  ā€¢ DNA amplification for diagnosis :
  ā€¢ Hepatitis C Virus- Detection of the virus by
    reverse transcriptase PCR confirms current
    infection and has a role in diagnosis and
    monitoring response to therapy. Development of
    Quantitative PCR allows quantification of HCV
    which is important in prognosis and monitoring
    of therapy.
  ā€¢ HIV : Newborns upto 18 months. Quantitation of
    HIV virus is widely used for prognosis and for
    evaluation of response to antiretroviral therapy

  2/7/2013                                         30
Molecular Biology contdā€¦


  Mycobacterium tuberculosis :
  ā€¢ In general the sensitivity of these assays for specimens
    for which the smear for acid fast bacilli (AFB) is positive
    is excellent (95-100%). It is lower for specimens that
    are AFB smear negative (50-80%).
  ā€¢ Currently, the amplification processes cannot replace
    the AFB smear because the latter is used to determine
    the level of infectivity of patients and in gauging the
    initial response to therapy.
  ā€¢ At present the role of amplification assays remains
    complimentary to microscopy and culture.

  2/7/2013                                                   31
Molecular Biology contdā€¦

  ā€¢ Sexually Transmitted Diseases : Amplification assays
    have been developed for Neisseria gonorrhoeae and
    Chlamydia trachomatis.
  ā€¢ Fungi : PCR assays for diagnosis of invasive
    Candidiasis and Aspergillosis have been developed.
  ā€¢ Detection of antimicrobial resistance : Molecular
    detection of the Mec-A gene is now considered a
    reference method for assessing Methicillin resistance
    in Staphylococci.
  ā€¢ Used in detection of resistance to rifampicin and
    isoniazid.
  2/7/2013                                             32
Molecular Biology contdā€¦


  Molecular Epidemiology :
  ā€¢ The techniques of molecular typing are useful
    not only in research settings but also in real
    life clinical and public health problems.
        ā€“ e.g. prediction of response to interferon therapy is
          important in patients with Hepatitis C virus
          infection.
  ā€¢ HCV genotypes 1b and 1a appear less likely to
    respond to interferons.

  2/7/2013                                                  33
Diagnostic Technologies in
Clinical Chemistry
ā€¢ Colorimetry : This is performed in a device that
  measures the absorbance of particular wavelengths
  of light by a specific solution.
ā€¢ Changeable optic filters are used in the colorimeter
  to select the wavelength of light which the solute
  absorbs the most, in order to maximize accuracy.
ā€¢ The usual wavelength range is from 400 to 700
  nanometres (nm). In modern colorimeters the
  filament lamp and filters may be replaced by several
  light-emitting diodes of different colors.
2/7/2013                                            34
Clinical Chemistry contdā€¦



  ā€¢ Applications in public health : Routine
    analytes like urea, creatinine, glucose, uric
    acid and also hemoglobin in hematology.
  ā€¢ Levels in blood of routine analytes forms the
    basis of the diagnosis of diabetes,
    hyperlipidemia, metabolic syndrome etc.



  2/7/2013                                      35
Clinical Chemistry contdā€¦


  Nephelometry :
  ā€¢ This is a method for measuring the concentration of a
    solution that contains particles that are too large for
    colorimetry/ absorption spectroscopy.
  ā€¢ When a collimated light beam strikes a particle in
    suspension, portions of the light are absorbed,
    reflected, scattered and transmitted.
  ā€¢ Nephelometry is the measurement of the light
    scattered by a particulate solution.
  ā€¢ Applications in public health : Measurement of
    antigen antibody complexes formed in immunoassays.
    Detection of IgG, IgM, IgA.

  2/7/2013                                               36
Clinical Chemistry contdā€¦


  Electrophoresis : Electrophoresis is the separation
    of charged compounds based on there electrical
    charge.
  ā€¢ Common support media include cellulose
    acetate, agarose and polyacrylamide gel.
  ā€¢ Once separation has occurred, the support
    medium is treated with dyes to stain and identify
    the separated fractions.
  ā€¢ To obtain a quantitative profile of the separated
    fractions, densitometry is performed on the
    stained support medium.
  2/7/2013                                          37
Clinical Chemistry contdā€¦


  Applications in public health :
  ā€¢ Separation of serum proteins :
     ā€“ Patterns of hypoproteinemia in malnutrition or gross
       loss of protein show decreases in all fractions with
       dramatic reduction in albumin.
     ā€“ Cirrhosis of the liver shows a specific pattern with
       severe reduction of albumin and increased
       immunoglobulins.
     ā€“ The pattern in monoclonal gammopathies (multiple
       myeloma) is very characteristic.
     ā€“ Separation of hemoglobins : This is important in the
       diagnosis of various abnormal hemoglobins like
       thalassemia, sickle cell anemia etc.

  2/7/2013                                               38
Clinical Chemistry contdā€¦


  Chromatography : Chromatography is a separation method based on
    the different interaction methods of the specimen compounds with
    the mobile phase and with the stationary phase, as the compounds
    travel through a support medium. The compounds interacting more
    strongly with the stationary phase are retained longer in the
    medium than those that favor the mobile phase.
  ā€¢ Gas chromatography : It is used for naturally volatile or those
    which can be easily converted to a volatile form.
  ā€¢ Liquid chromatography : It is used for too unstable or insufficiently
    volatile for gas chromatography.
  ā€¢ High Performance Liquid Chromatography : It is used for analyzing
    air and water pollutants, monitoring pesticide levels in the
    environment. Used In Sickle cell Disease diagnosis in infants.
    Detection of additives / chemicals in food.


  2/7/2013                                                             39
Clinical Chemistry contdā€¦



  ā€¢ Applications in public health : Used for
    determination of drugs and chemicals in body
    fluids (toxicology).
  ā€¢ Detection of additives / chemicals in food by
    High performance liquid chromatography.
  ā€¢ Environmental and water pollutants can be
    detected with accuracy through the use of
    various chromatographic procedures.

  2/7/2013                                      40
Clinical Chemistry contdā€¦


  Automation in clinical chemistry :
  ā€¢ Process a large volume of tests quickly. This is
    possible through the increased speed of testing.
  ā€¢ Most      automated      chemistry     analyzers,
    photometric methods of analysis such as
    colorimetry, spectrophotometry or nephelometry
    etc.
  ā€¢ Calculations, calibration curves and quality
    control are performed by the computers, thus
    reducing errors and providing more accurate
    results.
  2/7/2013                                          41
Clinical Chemistry contdā€¦


  ā€¢ The testing pathway in an analyser may be
    a) Sequential testing- Multiple tests analysed
       one after another on a single specimen.
    b) Batch testing- All samples are loaded at the
       same time and a single test is conducted on
       each sample.
    c) Parallel testing- More than one test is
       analysed concurrently on a given clinical
       specimen.
    d) Random access testing- Any test can be
       performed on any sample in any sequence.
  2/7/2013                                        42
Diagnostic Technologies in
                 Hematology
Automated Instruments :
ā€¢ These instruments have many different components to
  analyze different elements in the blood.
ā€¢ The cell counting components count the numbers and
  types of different cells within the blood. The results are
  printed out or sent to a computer for review.
ā€¢ It aspirate a very small amount of the specimen
  through narrow tubing. Within this tubing, there are
  sensors that count the number of cells going through it
  and can identify the type of cell; this is flow cytometry.
ā€¢ The two main sensors used are light detectors and
  electrical impedance.

2/7/2013                                                  43
Hematology contdā€¦


  ā€¢ One way the instrument can tell what type of
    blood cell is present is by size.
  ā€¢ Other       instruments       measure        different
    characteristics of the cells to categorize them.
  ā€¢ Because an automated cell counter samples and
    counts so many cells, the results are very precise.
  ā€¢ In addition to counting, measuring and analyzing
    red blood cells, white blood cells and platelets,
    automated hematology analyzers also measure
    the amount of hemoglobin in the blood and
    within each red blood cell.
  2/7/2013                                              44
Diagnostic Technologies in
Cytopathology
Exfoliative cytology : The microscopic examination of
  cells that have been shed from a lesion or have been
  recovered from a tissue for the diagnosis of disease.
Applications in public health
ā€¢ Cervical cytology : Exfoliative cytology from the
  uterine cervix is valuable in picking up premalignant
  lesions of the cervix.
ā€¢ It has an overall sensitivity of 80% and a specificity of
  99.4% for cytologic screening for cervical cancer.
ā€¢ The sensitivity was slightly lower for mild and
  moderate dysplasia (78.1%) and slightly higher for
  carcinoma in situ and severe dysplasia (81.4%) and
  invasive carcinoma (82.3%)
2/7/2013                                                 45
Cytopathology contdā€¦


  Fine Needle Aspiration Cytology :
  ā€¢ Fine Needle Aspiration biopsy is safer and less traumatic
     than an open surgical biopsy and significant complications
     are usually rare, depending on the body site.
  ā€¢ Common complications include bruising and soreness.
     There is also a risk that the cells taken do not enable a
     definitive diagnosis.
  ā€¢ The technique is suitable to evaluation of lesions/ nodules
     arising from breast, lymph nodes, thyroid, liver, kidney and
     soft tissue.
  ā€¢ The sensitivity and specificity varies according to the organ
     involved and also depends on the aspiration of a
     representativeness of the sample.

  2/7/2013                                                     46
Cytopathology contdā€¦



  Diagnostic Technologies in Histopathology
  ā€¢ The tissue is removed from the body and then placed
    in a fixative which stabilizes the tissues to prevent
    autolysis.
  ā€¢ The samples are transferred to a cassette, a container
    designed to allow reagents to freely act on the tissue
    inside. This process is known as tissue processing.
  ā€¢ The processed tissue is then taken out of the cassette
    and set in a mold. Through this process of embedding,
    additional paraffin is added to create a paraffin block.
  2/7/2013                                                47
Cytopathology contdā€¦


  ā€¢ The process of embedding then allows the
    sectioning of tissues into very thin (2 ā€“ 7
    micrometer) sections using a microtome.
  ā€¢ The microtome slices the tissue ready for
    microscopic examination. The slices are thinner
    than the average cell and are layered on a glass
    slide for staining.
  ā€¢ To see the tissue under a microscope, the
    sections are stained with one or more pigments.
  ā€¢ The most commonly used stain in histopathology
    is a combination of hematoxylin and eosin.
  2/7/2013                                         48
Cytopathology contdā€¦



  ā€¢ Applications in public health : Malignancies
    and also some non neoplastic conditions like
    tuberculosis and other infections.




  2/7/2013                                     49
Cytopathology contdā€¦


  ā€¢ Immunohistochemistry : Immunohistochemistry or
    IHC refers to the process of localizing proteins in cells
    of a tissue section exploiting the principle of antibodies
    binding specifically to antigens in biological tissues.
  ā€¢ Immunohistochemical staining is widely used in the
    diagnosis of abnormal cells such as those found in
    cancerous tumors.
  ā€¢ Applications in public health : Recognition of specific
    molecular markers like S100, Cytokeratin etc. helps in
    identifying the origin and hence the diagnosis of
    various tumours and conditions.

  2/7/2013                                                  50
Radiological Techniques

X-ray : For detection of pathology of the skeletal system
  and some disease processes in soft tissue, e.g.
  pneumonia, lung cancer or pulmonary edema and the
  abdominal X-ray, which can detect ileus (blockage of
  the intestine), free air (from visceral perforations) and
  free fluid (in ascites).
Applications in public health : Chest x ray was the
  backbone of National Tuberculosis Control Programme
  in 1962.
ā€¢ However it is included in the diagnostic algorithm of
  RNTCP where no sputum sample is positive/ two
  negative smears in a symptomatic patient.
ā€¢ In occupational health the ILO criterion for diagnosis of
  pneumoconiosis uses findings of x ray chest only.
2/7/2013                                                 51
Radiology contdā€¦


  ā€¢ DEXASCAN : DEXA Scan stands for Dual Energy X-
    Ray Absorptiometry. Two different types of X- ray
    scan the body and the images are subtracted by
    the computer giving a very accurate estimation of
    bone density/body fat.
  ā€¢ It is used to detect the presence of osteoporosis
    in men and women with particular risk factors,
    screen for osteoporosis, particularly in women
    making decisions about hormone replacement
    therapy at menopause.

  2/7/2013                                          52
Radiology contdā€¦



  ā€¢ Angiography : It is the study of blood vessels
    with a contrast medium.
  ā€¢ It has a great role in study of coronaries to
    detect the status of these vessels and decide
    the treatment modality for the patient.
  ā€¢ With the epidemic of coronary artery disease
    angiography will go a long way in the final
    management of the patients.

  2/7/2013                                       53
Radiology contdā€¦

 Mammography :
 ā€¢ Mammography is the process of using low-dose
   X-rays, high contrast, high-resolution film and an
   X-ray system designed specifically for imaging the
   breasts.
 ā€¢ The goal of mammography is the early detection
   of breast cancer, typically through detection of
   characteristic masses and/or micro calcifications.
 ā€¢ Mammography has been shown to reduce
   mortality from breast cancer.

  2/7/2013                                          54
Radiology contdā€¦

  ā€¢ USG:-
        ā€“ Obstetric use
        ā€“ Cardiac scan
        ā€“ Renal, liver and gallbladder scans.
        ā€“ Musculo-skeletal imaging of muscles, ligaments
          and tendons,
        ā€“ Eye scans
        ā€“ Testicle, thyroid, salivary glands and lymph nodes
          scan.
        ā€“ To guide interventional procedures (FNAC)
  2/7/2013                                                55
Radiology contdā€¦

  ā€¢ Applications in Public Health : This technique in
    antenatal cases is practiced to
        ā€“    Date the pregnancy
        ā€“    Confirm fetal viability,
        ā€“    Determine location of fetus,
        ā€“    Intrauterine vs ectopic,
        ā€“    Location of the placenta.
        ā€“    Number of fetuses
        ā€“    Major physical abnormalities,
        ā€“    Assess fetal growth
        ā€“    Fetal movement and heartbeat.
  ā€¢ Ultrasound is also increasingly being used in trauma
    and first aid cases, with emergency ultrasound
    becoming a stable of most emergency response teams.
  2/7/2013                                            56
Radiology contdā€¦


  CT scan :
  ā€¢ CT scan works on the principle that cross
    sectional slices of the body are produced using X
    rays, followed by processing by the computers to
    study detailed anatomy of the specific location or
    slice.
  ā€¢ The modification of conventional CT is spiral
    where the X-ray tube/ detector combination
    rotates continuously around the patient creating
    a spiral and helps in generating high quality
    images.

  2/7/2013                                          57
Radiology contdā€¦


  ā€¢ Magnetic Resonance Imaging : Charged spinning
    particles such a protons behave like tiny bar magnets
    and are used to produce image of biological tissue.
  ā€¢ It does not use ionizing radiation hence is safe with no
    known biological hazards.
  ā€¢ It provides excellent tissue contrast with good
    resolution.
  ā€¢ Images can be obtained in any plane (Multiplanar
    Imaging) and does not produce artifacts due to bone
    and is an ideal imaging modality for spine, posterior
    fossa and musculoskeletal system.

  2/7/2013                                                58
Radiology contdā€¦


  Radionuclide scan :
  ā€¢ A radionuclide scan is a way of imaging bones, organs and
    other parts of the body by using a small dose of a
    radioactive chemical.
  ā€¢ Cells which are most ā€˜activeā€™ in the target tissue or organ
    will take up more of the radionuclide. So, active parts of the
    tissue will emit more gamma rays than less active or
    inactive parts.
  ā€¢ The gamma rays which are emitted from inside the body
    are detected by the gamma camera, are converted into an
    electrical signal and sent to a computer.
  ā€¢ The computer builds a picture by converting the differing
    intensities of radioactivity emitted into different colours or
    shades of grey.
  2/7/2013                                                      59
Radiology contdā€¦



  ā€¢ Public Health Importance
        ā€“ Bone scan : Used to detect areas of bone where
          there is cancer, infection or damage.
        ā€“ Kidney scan : Used to detect scars on the kidney
          and how well urine drains from the kidney to the
          bladder.
        ā€“ Lung Perfusion Scan : Detects pulmonary
          embolus.


  2/7/2013                                              60
CT scan, its modifications and their uses in
               public health




2/7/2013                                   61
MRI : Clinical uses




2/7/2013                         62
IPHS Standards : Suggested
Availability of radiological studies




2/7/2013                           63
Electrophysical Technologies

ā€¢ Various functional disturbances sometimes detectable
  only via analysis of electric potentials produced by
  some organs (tissues) as a manifestation of their
  function (ECG, EEG, EMG, ERG and others).
ā€¢ Evaluation of these signals is performed either in the
  ā€œtime domainā€ (detection of particular specific peaks
  and wave complexes - ā€œgrapho-elementsā€ - and
  description of their time and amplitude characteristics)
  or in the ā€œfrequency domainā€ (frequency spectrum
  characteristics).
ā€¢ At present this is done almost exclusively with the use
  of computers.

2/7/2013                                                64
Electrophysiology contdā€¦


  Electroencephalography :
  ā€¢ This is the neurophysiologic measurement of the electrical
     activity of the brain by recording from electrodes placed on
     the scalp or in special cases, subdurally or in the cerebral
     cortex. It represent an electrical signal (postsynaptic
     potentials) from a large number of neurons.
  ā€¢ Each electrode is connected to an input of a differential
     amplifier, which amplifies the voltage between them. The
     resulting voltage signal is filtered which is shown on paper
     or displayed on a computer screen. Uses in
        ā€“ Epilepsy and syncope
        ā€“ Sleep disorders
        ā€“ Coma and brain death

  2/7/2013                                                     65
Electrophysiology contdā€¦


  Electrocardiography : An electrocardiogram records the
     electrical activity of the heart over time.
  ā€¢ Electrodes on different sides of the heart measure the
     activity of different parts of the heart muscle.
  ā€¢ An ECG displays the overall rhythm of the heart and
     weaknesses in different parts of the heart muscle.
  ā€¢ Applications in public health : It is the best way to measure
     and diagnose abnormal rhythms of the heart in
      ā€“ Damage to the conductive tissue carrying electrical
        signals,
      ā€“ High or low potassium.
      ā€“ Myocardial infarction (MI)

  2/7/2013                                                     66
Electrophysiology contdā€¦



  Electromyography :
  ā€¢ Electromyography (EMG) is a technique for
    evaluating and recording the activation signal of
    muscles.
  ā€¢ An electromyograph detects the electrical
    potential generated by muscle cells when these
    cells contract and also when the cells are at rest.
  ā€¢ EMG is used to diagnose two general categories
    of disease : neuropathies and myopathies.
  2/7/2013                                           67
ENDOSCopy

ā€¢ Different types of endoscopies are performed for diagnostic
  purposes of carcinoma and other pathologies. These may
  be rigid or fibreoptic.
ā€¢ Direct:-
      ā€“    Laryngoscopy
      ā€“    Broncoscopy
      ā€“    Colonoscopy
      ā€“    Gastroscopy
      ā€“    Nasopharyngoscopy
      ā€“    Sigmoidoscopy
      ā€“    Hysteroscopy
ā€¢ Indirect:-
      ā€“ Laryngoscopy

2/7/2013                                                   68
References

ā€¢ Rajvir Balwar, Text book Public Health and Community Medicine,
  1st Edition, AFMC Pune in collabration with WHO, Dept Of
  Commnity Medicine, 2008.
ā€¢ Indian Public Health Standards (IPHS) Guidelines for Sub-
  District/Sub-Divisional Hospitals (31 to 100 Bedded) Revised 2012 ,
  DGHS, MOHFW, Govt Of India, June 2012.
ā€¢ Indian Public Health Standards (IPHS) Guidelines for District
  Hospitals (101 to 500 Bedded) Revised 2012, DGHS, MOHFW, Govt
  Of India, June 2012.
ā€¢ http://www.biomerieux.com/en/advanced-technologies
ā€¢ World Health Organization, Laboratory Support to Emerging
  Diseases. World Health Organization Regional Office for South-East
  Asia New Delhi, 2008.
ā€¢ www.whoindia.org/LinkFiles/Public_Health_Laboratory_Networkin
  g_SEARO_recommendations _on_networking_of_PHL

2/7/2013                                                           69
ā€¢ The Diagnostics Innovation Map: Medical Diagnostics for the Unmet
  Needs of the Developing World, WHO BIO Ventures for Global
  Health,2010.
ā€¢ Estimating the Global Health Impact of Improved Diagnostic Tools
  for the Developing World. Santa Monica, Rand Institute,2007.
ā€¢ Sen J, Belli A "S100B in neuropathologic states: the CRP of the
  brain?". J. Neurosci. Res.2007; 85 (7): 1373ā€“80.
ā€¢ http://mohfw.nic.in/NRHM/iphs.htm
ā€¢ http://testequipment.globalspec.com/LearnMore/Labware_Scientif
  ic_Instruments/Analytical_Instruments/Chromatographs/HPLC
ā€¢ http://www.google.co.in/imgres?um/diagnostic%technologies/php.




2/7/2013                                                         70
2/7/2013   71

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Advanced Diagnostic Technologies in Public Health

  • 1. Advanced Diagnostic Technologies in Public Health Dr Abhay Dhanorkar 2/7/2013 1
  • 2. Scope ā€¢ Introduction ā€¢ IPHS standards for availability of laboratory services at various hospitals. ā€¢ Classification of diagnostics tests ā€¢ Immunopathology ā€¢ Molecular Biology ā€¢ Clinical Chemistry ā€¢ Haematology ā€¢ Cytopathology ā€¢ Histopathology ā€¢ Radiology ā€¢ Electrophysiology ā€¢ Endoscopy ā€¢ Summary 2/7/2013 2
  • 3. Introduction ā€¢ A diagnostic test is any kind of medical test performed to aid in the diagnosis or detection of disease. ā€¢ It may be used to diagnose diseases or measure the progress or recovery from disease or confirm that a person is free from disease. ā€¢ Some medical tests are parts of a simple physical examination which require only simple tools in the hands of a skilled practitioner and can be performed in an office environment. 2/7/2013 3
  • 4. Introduction contdā€¦ā€¦ ā€¢ Some other tests require elaborate equipment used by medical technologists or the use of a sterile operating theatre environment. ā€¢ Some tests require samples of tissue or body fluids to be sent off to a pathology lab for further analysis. ā€¢ Some simple chemical tests such as urine pH, can be measured directly in the physicians office. 2/7/2013 4
  • 5. Introduction contdā€¦ā€¦ ā€¢ However, with advancements in health care, a large array of diagnostic tests have become available. Various modern (and often, expensive) diagnostic modalities are often routinely used in medical world. ā€¢ According to IPHS standards stated as a part of NRHM, the some laboratory facilities should be available at various levels. 2/7/2013 5
  • 6. IPHS standards for availability of laboratory services at 31 to 50 bedded hospitals 2/7/2013 6
  • 7. For 51-100 Bedded (All as in 31-50 bedded with the following additional facilities) 2/7/2013 7
  • 8. For 101-200 Bedded (All as in 51-100 bedded with the following additional facilities) Speciality Diagnostic Services / Tests Colonoscopy, Bronchoscopy, Arthroscopy, Laparoscopy Endoscopy (Diagnostic), Colposcopy, Hysteroscopy Serology RA factor test CPK, Lepto spirosis (Rapid test / ELISA) HSG, Ultrasonography Colour Doppler, Spiral CT scan, Radiology MRI 0.5 TESSLA Biochemistry Glycosylated Hemoglobin, Blood gas analysis 2/7/2013 8
  • 9. For 201-300 Bedded (All as in 101-200 bedded with the following additional facilities) 2/7/2013 9
  • 10. For >300 Bedded (All as in 201-300 bedded with the following additional facilities) Speciality Diagnostic Services Angiography, Multislice CT, Multislice Radiology MRI Bone Marrow Biopsy,Pleural Biopsy, Biopsy Pericardial tapping Gastroscopy (Oesophagus, stomach, Endoscopic Specialised Procedures deudenum), Sigmoid, Laproscopy and and Diagnostic Colonoscopy, Bronchoscopy and Foreign Body Removal 2/7/2013 10
  • 11. Classification ā€¢ The various modern diagnostic technologies can be classified as ā€“ Immunopathology ā€“ Molecular Biology ā€“ Clinical Chemistry ā€“ Haematology ā€“ Cytopathology ā€“ Histopathology ā€“ Radiology ā€“ Electrophysiology ā€“ Endoscopy 2/7/2013 11
  • 12. Diagnostic Technologies in Immunopathology ā€¢ Antibodies are produced in response to antigenic stimulation. ā€¢ Immunoassays (antigen antibody reactions) can be used for detection of either antigens or antibodies. For detecting antigens, the corresponding specific antibody should be used as one of the reagents and vice versa. 2/7/2013 12
  • 13. Immunopathology contdā€¦ The technologies available are ā€“ ā€¢ Precipitation immunoassays : It is based on the occurrence of precipitation when large complexes of antigens and antibodies combine to form an insoluble lattice. Ex. :- Widal test and the Weil-Felix test. ā€¢ Particle immunoassay : Here specific antigens are coated onto a particle and on reaction with antibody under test, the agglutination is made more visible. Reverse agglutination where the antibody is coated onto the particle is also performed. The particle used may be RBC, latex or gelatin. Ex. :- 1) TPHA (Treponema pallidum Hemagglutination). 2) Reverse passive hemagglutination (RPHA) for hepatitis B surface antigen (HBsAg). 3) Latex agglutination is used in measurement of hCG for qualitative pregnancy tests 2/7/2013 13
  • 14. Immunopathology contdā€¦ Radioimmunoassay : ā€¢ Radioimmunoassay (RIA) is used to test antigens (e.g.hormone levels in the blood). ā€¢ It is extremely sensitive and extremely specific, but requires a sophisticated apparatus and is costly. It also requires special precautions, since radioactive substances are used. ā€¢ Therefore, today it has been largely supplanted by the ELISA method, where the antigen-antibody reaction is measured using colorometric signals instead of a radioactive signal. ā€¢ To perform a radioimmunoassay, a known quantity of an antigen is made radioactive, frequently by labeling it with gamma-radioactive isotopes of iodine attached to thyrosine. 2/7/2013 14
  • 15. Immunopathology contdā€¦ ā€¢ This radiolabeled antigen is then mixed with a known amount of antibody for that antigen and as a result, the two chemically bind to one another. ā€¢ Then, a sample of serum from a patient containing an unknown quantity of that same antigen is added. This causes the unlabeled (or ā€œcoldā€) antigen from the serum to compete with the radiolabeled antigen for antibody binding sites. Applications in public health : Ex. Thyroid hormone levels in goiter endemic areas. 2/7/2013 15
  • 16. Immunopathology contdā€¦ Enzyme Linked Immunosorbent Assay (ELISA) : ā€¢ This is a technique used mainly in immunology to detect the presence of an antibody or an antigen in a sample. ā€¢ In ELISA an unknown amount of antigen is affixed to a surface and then a specific antibody is washed over the surface so that it can bind to the antigen. ā€¢ This antibody is linked to an enzyme and in the final step a substance is added that the enzyme can convert to some detectable signal. 2/7/2013 16
  • 17. Immunopathology contdā€¦ Applications in public health : ā€“ HIV test or West Nile Virus. ELISA can be used for detecting tumour markers for certain cancers, e.g. Prostate Specific Antigen (PSA) in carcinoma prostate. 2/7/2013 17
  • 18. Immunopathology contdā€¦ Fluorescent immunoassay : ā€¢ When fluorescent molecules are irradiated with light at appropriate wavelengths, an electron in the ground state is transited into the excited state. As the electron returns to the ground state, physical energy is released in the form of a photon which is detectable. Applications in public health : ā€¢ For detecting antigens in tissue sections in kidney biopsies. ā€¢ Use of Auramine rhodamine stain in detection of tubercle bacilli in smears shown to have better sensitivity than the conventional ZN stain. ā€¢ To detect the concentration of drugs, hormones and proteins and polypeptides. 2/7/2013 18
  • 19. Diagnostic Technologies in Molecular Biology Diagnostic Technologies in Molecular Biology ā€¢ Molecular biology is the study of biology at a molecular level. ā€¢ Molecular biology chiefly concerns itself with understanding the interactions between the various systems of a cell, including the interactions between DNA, RNA and protein biosynthesis and learning how these interactions are regulated. ā€¢ The tools of molecular biology is extremely useful in diagnosis, therapy, epidemiologic investigations and infection control. 2/7/2013 19
  • 20. Molecular Biology contdā€¦ ā€¢ Ease of performance, reproducibility, sensitivity and specificity of molecular tests are important, cost and potential contribution to patient care are also of concern. ā€¢ Molecular methods may be an improvement over conventional microbiologic testing as they are most practical and useful application is in detecting and identifying infectious agents for which routine growth-based culture and microscopy methods may not be adequate. 2/7/2013 20
  • 21. Molecular Biology contdā€¦ Gel electrophoresis : ā€¢ It is one of the principal tools of molecular biology. The basic principle is that DNA, RNA and proteins can all be separated by means of an electric field. ā€¢ In agarose gel electrophoresis, DNA and RNA can be separated on the basis of size by running the DNA through an agarose gel. ā€¢ Proteins can be separated on the basis of size by using an SDS-PAGE gel, or on the basis of size and their electric charge by using 2D gel electrophoresis. 2/7/2013 21
  • 22. Molecular Biology contdā€¦ Hybridisation assays : ā€¢ When the hybridization reaction is used to analyse the nucleic acid content of an unknown sample, the process is known as a hybridization assay. ā€¢ The property of complimentary base pairing allows fragments of known composition (the probes) to interrogate an unknown for the presence of matching (complimentary) sequences. ā€¢ The detection of the hybrids can be done a variety of technologies including radioisotope labels, fluorochrome based detection and enzyme based systems. 2/7/2013 22
  • 23. Molecular Biology contdā€¦ ā€¢ Liquid phase hybridization : When both sample and probe are in solution. ā€¢ Solid support hybridization : In these assays, hybridization occurs in a biphasic environment, a solid phase (usually sample) and a liquid phase (usually probe). ā€¢ Southern and Northern hybridization assays : These combine electrophoretic separation of test nucleic acids with transfer to a solid support and subsequent hybridization. 2/7/2013 23
  • 24. Molecular Biology contdā€¦ ā€¢ The original procedure was described by EM Southern and the test nucleic acid was DNA. When RNA is the nucleic acid under test, the technique is called northern blotting by analogy. ā€¢ In situ hybridization : This is the detection of specific genetic information within a morphological context (intact tissue, cells). 2/7/2013 24
  • 25. Molecular Biology contdā€¦ Amplification technology : ā€¢ Amplification systems are enzyme based processes in which a single enzyme or multiple enzymes synthesize copies of target nucleic acid. ā€¢ All result in production of billions of copies of the amplified product in a few hours. ā€¢ These techniques are subject to contamination from product molecules of previous amplifications and hence false positivity is high. ā€¢ However, special lab design, practices and workflow have helped to reduce false positives to an acceptable range. 2/7/2013 25
  • 26. Molecular Biology contdā€¦ Polymerase Chain Reaction (PCR) ā€¢ It is a simple in vitro chemical reaction that permits synthesis of large quantities of nucleic acid. ā€¢ This is brought about by heating the reaction mixture to separate the strands of the DNA, then cooled to permit the primers to anneal to the target DNA in a sequence specific manner. The DNA polymerase then initiates extension from end. ā€¢ Thus the whole process is carried out in a programmable thermocycler which controls the temperatures at which various steps occur. 2/7/2013 26
  • 27. Molecular Biology contdā€¦ ā€¢ Reverse transcriptase PCR (RT-PCR) : PCR was initially described to amplify DNA. RT-PCR was developed to amplify RNA targets ā€¢ Nested PCR : This variation increases the sensitivity and specificity of PCR. The products of the first round of amplification are subject to second round of amplification with a second set of primers. ā€¢ Real time PCR : Here the target amplification and detection occur simultaneously. ā€¢ The computer software supporting the thermocycler monitors the data throughout the PCR at every cycle and generates a plot. 2/7/2013 27
  • 28. Molecular Biology contdā€¦ Applications of molecular biology in Public Health ā€¢ Pathogen detection by hybridisation : The DNA of the infectious agent is detected directly in the clinical specimens by DNA probes. 2/7/2013 28
  • 29. Molecular Biology contdā€¦ ā€¢ DNA probes for culture identification : Probes are available for various organisms like Mycobacterium tuberculosis, Chlamydia trachomatis, Neisseria gonorrhoeae, Human papilloma virus, Hepatitis C virus, Cytomegalovirus and Herpes simplex virus. ā€¢ This is especially important for slow growing organisms like Mycobacteria, fungi etc. Identification of cultured Mycobacteria by conventional methods is slow and time consuming. ā€¢ The use of probes for this purpose permits identification from cultures within one working day and sensitivity (95.4%) and specificity (99%) are excellent. 2/7/2013 29
  • 30. Molecular Biology contdā€¦ ā€¢ DNA amplification for diagnosis : ā€¢ Hepatitis C Virus- Detection of the virus by reverse transcriptase PCR confirms current infection and has a role in diagnosis and monitoring response to therapy. Development of Quantitative PCR allows quantification of HCV which is important in prognosis and monitoring of therapy. ā€¢ HIV : Newborns upto 18 months. Quantitation of HIV virus is widely used for prognosis and for evaluation of response to antiretroviral therapy 2/7/2013 30
  • 31. Molecular Biology contdā€¦ Mycobacterium tuberculosis : ā€¢ In general the sensitivity of these assays for specimens for which the smear for acid fast bacilli (AFB) is positive is excellent (95-100%). It is lower for specimens that are AFB smear negative (50-80%). ā€¢ Currently, the amplification processes cannot replace the AFB smear because the latter is used to determine the level of infectivity of patients and in gauging the initial response to therapy. ā€¢ At present the role of amplification assays remains complimentary to microscopy and culture. 2/7/2013 31
  • 32. Molecular Biology contdā€¦ ā€¢ Sexually Transmitted Diseases : Amplification assays have been developed for Neisseria gonorrhoeae and Chlamydia trachomatis. ā€¢ Fungi : PCR assays for diagnosis of invasive Candidiasis and Aspergillosis have been developed. ā€¢ Detection of antimicrobial resistance : Molecular detection of the Mec-A gene is now considered a reference method for assessing Methicillin resistance in Staphylococci. ā€¢ Used in detection of resistance to rifampicin and isoniazid. 2/7/2013 32
  • 33. Molecular Biology contdā€¦ Molecular Epidemiology : ā€¢ The techniques of molecular typing are useful not only in research settings but also in real life clinical and public health problems. ā€“ e.g. prediction of response to interferon therapy is important in patients with Hepatitis C virus infection. ā€¢ HCV genotypes 1b and 1a appear less likely to respond to interferons. 2/7/2013 33
  • 34. Diagnostic Technologies in Clinical Chemistry ā€¢ Colorimetry : This is performed in a device that measures the absorbance of particular wavelengths of light by a specific solution. ā€¢ Changeable optic filters are used in the colorimeter to select the wavelength of light which the solute absorbs the most, in order to maximize accuracy. ā€¢ The usual wavelength range is from 400 to 700 nanometres (nm). In modern colorimeters the filament lamp and filters may be replaced by several light-emitting diodes of different colors. 2/7/2013 34
  • 35. Clinical Chemistry contdā€¦ ā€¢ Applications in public health : Routine analytes like urea, creatinine, glucose, uric acid and also hemoglobin in hematology. ā€¢ Levels in blood of routine analytes forms the basis of the diagnosis of diabetes, hyperlipidemia, metabolic syndrome etc. 2/7/2013 35
  • 36. Clinical Chemistry contdā€¦ Nephelometry : ā€¢ This is a method for measuring the concentration of a solution that contains particles that are too large for colorimetry/ absorption spectroscopy. ā€¢ When a collimated light beam strikes a particle in suspension, portions of the light are absorbed, reflected, scattered and transmitted. ā€¢ Nephelometry is the measurement of the light scattered by a particulate solution. ā€¢ Applications in public health : Measurement of antigen antibody complexes formed in immunoassays. Detection of IgG, IgM, IgA. 2/7/2013 36
  • 37. Clinical Chemistry contdā€¦ Electrophoresis : Electrophoresis is the separation of charged compounds based on there electrical charge. ā€¢ Common support media include cellulose acetate, agarose and polyacrylamide gel. ā€¢ Once separation has occurred, the support medium is treated with dyes to stain and identify the separated fractions. ā€¢ To obtain a quantitative profile of the separated fractions, densitometry is performed on the stained support medium. 2/7/2013 37
  • 38. Clinical Chemistry contdā€¦ Applications in public health : ā€¢ Separation of serum proteins : ā€“ Patterns of hypoproteinemia in malnutrition or gross loss of protein show decreases in all fractions with dramatic reduction in albumin. ā€“ Cirrhosis of the liver shows a specific pattern with severe reduction of albumin and increased immunoglobulins. ā€“ The pattern in monoclonal gammopathies (multiple myeloma) is very characteristic. ā€“ Separation of hemoglobins : This is important in the diagnosis of various abnormal hemoglobins like thalassemia, sickle cell anemia etc. 2/7/2013 38
  • 39. Clinical Chemistry contdā€¦ Chromatography : Chromatography is a separation method based on the different interaction methods of the specimen compounds with the mobile phase and with the stationary phase, as the compounds travel through a support medium. The compounds interacting more strongly with the stationary phase are retained longer in the medium than those that favor the mobile phase. ā€¢ Gas chromatography : It is used for naturally volatile or those which can be easily converted to a volatile form. ā€¢ Liquid chromatography : It is used for too unstable or insufficiently volatile for gas chromatography. ā€¢ High Performance Liquid Chromatography : It is used for analyzing air and water pollutants, monitoring pesticide levels in the environment. Used In Sickle cell Disease diagnosis in infants. Detection of additives / chemicals in food. 2/7/2013 39
  • 40. Clinical Chemistry contdā€¦ ā€¢ Applications in public health : Used for determination of drugs and chemicals in body fluids (toxicology). ā€¢ Detection of additives / chemicals in food by High performance liquid chromatography. ā€¢ Environmental and water pollutants can be detected with accuracy through the use of various chromatographic procedures. 2/7/2013 40
  • 41. Clinical Chemistry contdā€¦ Automation in clinical chemistry : ā€¢ Process a large volume of tests quickly. This is possible through the increased speed of testing. ā€¢ Most automated chemistry analyzers, photometric methods of analysis such as colorimetry, spectrophotometry or nephelometry etc. ā€¢ Calculations, calibration curves and quality control are performed by the computers, thus reducing errors and providing more accurate results. 2/7/2013 41
  • 42. Clinical Chemistry contdā€¦ ā€¢ The testing pathway in an analyser may be a) Sequential testing- Multiple tests analysed one after another on a single specimen. b) Batch testing- All samples are loaded at the same time and a single test is conducted on each sample. c) Parallel testing- More than one test is analysed concurrently on a given clinical specimen. d) Random access testing- Any test can be performed on any sample in any sequence. 2/7/2013 42
  • 43. Diagnostic Technologies in Hematology Automated Instruments : ā€¢ These instruments have many different components to analyze different elements in the blood. ā€¢ The cell counting components count the numbers and types of different cells within the blood. The results are printed out or sent to a computer for review. ā€¢ It aspirate a very small amount of the specimen through narrow tubing. Within this tubing, there are sensors that count the number of cells going through it and can identify the type of cell; this is flow cytometry. ā€¢ The two main sensors used are light detectors and electrical impedance. 2/7/2013 43
  • 44. Hematology contdā€¦ ā€¢ One way the instrument can tell what type of blood cell is present is by size. ā€¢ Other instruments measure different characteristics of the cells to categorize them. ā€¢ Because an automated cell counter samples and counts so many cells, the results are very precise. ā€¢ In addition to counting, measuring and analyzing red blood cells, white blood cells and platelets, automated hematology analyzers also measure the amount of hemoglobin in the blood and within each red blood cell. 2/7/2013 44
  • 45. Diagnostic Technologies in Cytopathology Exfoliative cytology : The microscopic examination of cells that have been shed from a lesion or have been recovered from a tissue for the diagnosis of disease. Applications in public health ā€¢ Cervical cytology : Exfoliative cytology from the uterine cervix is valuable in picking up premalignant lesions of the cervix. ā€¢ It has an overall sensitivity of 80% and a specificity of 99.4% for cytologic screening for cervical cancer. ā€¢ The sensitivity was slightly lower for mild and moderate dysplasia (78.1%) and slightly higher for carcinoma in situ and severe dysplasia (81.4%) and invasive carcinoma (82.3%) 2/7/2013 45
  • 46. Cytopathology contdā€¦ Fine Needle Aspiration Cytology : ā€¢ Fine Needle Aspiration biopsy is safer and less traumatic than an open surgical biopsy and significant complications are usually rare, depending on the body site. ā€¢ Common complications include bruising and soreness. There is also a risk that the cells taken do not enable a definitive diagnosis. ā€¢ The technique is suitable to evaluation of lesions/ nodules arising from breast, lymph nodes, thyroid, liver, kidney and soft tissue. ā€¢ The sensitivity and specificity varies according to the organ involved and also depends on the aspiration of a representativeness of the sample. 2/7/2013 46
  • 47. Cytopathology contdā€¦ Diagnostic Technologies in Histopathology ā€¢ The tissue is removed from the body and then placed in a fixative which stabilizes the tissues to prevent autolysis. ā€¢ The samples are transferred to a cassette, a container designed to allow reagents to freely act on the tissue inside. This process is known as tissue processing. ā€¢ The processed tissue is then taken out of the cassette and set in a mold. Through this process of embedding, additional paraffin is added to create a paraffin block. 2/7/2013 47
  • 48. Cytopathology contdā€¦ ā€¢ The process of embedding then allows the sectioning of tissues into very thin (2 ā€“ 7 micrometer) sections using a microtome. ā€¢ The microtome slices the tissue ready for microscopic examination. The slices are thinner than the average cell and are layered on a glass slide for staining. ā€¢ To see the tissue under a microscope, the sections are stained with one or more pigments. ā€¢ The most commonly used stain in histopathology is a combination of hematoxylin and eosin. 2/7/2013 48
  • 49. Cytopathology contdā€¦ ā€¢ Applications in public health : Malignancies and also some non neoplastic conditions like tuberculosis and other infections. 2/7/2013 49
  • 50. Cytopathology contdā€¦ ā€¢ Immunohistochemistry : Immunohistochemistry or IHC refers to the process of localizing proteins in cells of a tissue section exploiting the principle of antibodies binding specifically to antigens in biological tissues. ā€¢ Immunohistochemical staining is widely used in the diagnosis of abnormal cells such as those found in cancerous tumors. ā€¢ Applications in public health : Recognition of specific molecular markers like S100, Cytokeratin etc. helps in identifying the origin and hence the diagnosis of various tumours and conditions. 2/7/2013 50
  • 51. Radiological Techniques X-ray : For detection of pathology of the skeletal system and some disease processes in soft tissue, e.g. pneumonia, lung cancer or pulmonary edema and the abdominal X-ray, which can detect ileus (blockage of the intestine), free air (from visceral perforations) and free fluid (in ascites). Applications in public health : Chest x ray was the backbone of National Tuberculosis Control Programme in 1962. ā€¢ However it is included in the diagnostic algorithm of RNTCP where no sputum sample is positive/ two negative smears in a symptomatic patient. ā€¢ In occupational health the ILO criterion for diagnosis of pneumoconiosis uses findings of x ray chest only. 2/7/2013 51
  • 52. Radiology contdā€¦ ā€¢ DEXASCAN : DEXA Scan stands for Dual Energy X- Ray Absorptiometry. Two different types of X- ray scan the body and the images are subtracted by the computer giving a very accurate estimation of bone density/body fat. ā€¢ It is used to detect the presence of osteoporosis in men and women with particular risk factors, screen for osteoporosis, particularly in women making decisions about hormone replacement therapy at menopause. 2/7/2013 52
  • 53. Radiology contdā€¦ ā€¢ Angiography : It is the study of blood vessels with a contrast medium. ā€¢ It has a great role in study of coronaries to detect the status of these vessels and decide the treatment modality for the patient. ā€¢ With the epidemic of coronary artery disease angiography will go a long way in the final management of the patients. 2/7/2013 53
  • 54. Radiology contdā€¦ Mammography : ā€¢ Mammography is the process of using low-dose X-rays, high contrast, high-resolution film and an X-ray system designed specifically for imaging the breasts. ā€¢ The goal of mammography is the early detection of breast cancer, typically through detection of characteristic masses and/or micro calcifications. ā€¢ Mammography has been shown to reduce mortality from breast cancer. 2/7/2013 54
  • 55. Radiology contdā€¦ ā€¢ USG:- ā€“ Obstetric use ā€“ Cardiac scan ā€“ Renal, liver and gallbladder scans. ā€“ Musculo-skeletal imaging of muscles, ligaments and tendons, ā€“ Eye scans ā€“ Testicle, thyroid, salivary glands and lymph nodes scan. ā€“ To guide interventional procedures (FNAC) 2/7/2013 55
  • 56. Radiology contdā€¦ ā€¢ Applications in Public Health : This technique in antenatal cases is practiced to ā€“ Date the pregnancy ā€“ Confirm fetal viability, ā€“ Determine location of fetus, ā€“ Intrauterine vs ectopic, ā€“ Location of the placenta. ā€“ Number of fetuses ā€“ Major physical abnormalities, ā€“ Assess fetal growth ā€“ Fetal movement and heartbeat. ā€¢ Ultrasound is also increasingly being used in trauma and first aid cases, with emergency ultrasound becoming a stable of most emergency response teams. 2/7/2013 56
  • 57. Radiology contdā€¦ CT scan : ā€¢ CT scan works on the principle that cross sectional slices of the body are produced using X rays, followed by processing by the computers to study detailed anatomy of the specific location or slice. ā€¢ The modification of conventional CT is spiral where the X-ray tube/ detector combination rotates continuously around the patient creating a spiral and helps in generating high quality images. 2/7/2013 57
  • 58. Radiology contdā€¦ ā€¢ Magnetic Resonance Imaging : Charged spinning particles such a protons behave like tiny bar magnets and are used to produce image of biological tissue. ā€¢ It does not use ionizing radiation hence is safe with no known biological hazards. ā€¢ It provides excellent tissue contrast with good resolution. ā€¢ Images can be obtained in any plane (Multiplanar Imaging) and does not produce artifacts due to bone and is an ideal imaging modality for spine, posterior fossa and musculoskeletal system. 2/7/2013 58
  • 59. Radiology contdā€¦ Radionuclide scan : ā€¢ A radionuclide scan is a way of imaging bones, organs and other parts of the body by using a small dose of a radioactive chemical. ā€¢ Cells which are most ā€˜activeā€™ in the target tissue or organ will take up more of the radionuclide. So, active parts of the tissue will emit more gamma rays than less active or inactive parts. ā€¢ The gamma rays which are emitted from inside the body are detected by the gamma camera, are converted into an electrical signal and sent to a computer. ā€¢ The computer builds a picture by converting the differing intensities of radioactivity emitted into different colours or shades of grey. 2/7/2013 59
  • 60. Radiology contdā€¦ ā€¢ Public Health Importance ā€“ Bone scan : Used to detect areas of bone where there is cancer, infection or damage. ā€“ Kidney scan : Used to detect scars on the kidney and how well urine drains from the kidney to the bladder. ā€“ Lung Perfusion Scan : Detects pulmonary embolus. 2/7/2013 60
  • 61. CT scan, its modifications and their uses in public health 2/7/2013 61
  • 62. MRI : Clinical uses 2/7/2013 62
  • 63. IPHS Standards : Suggested Availability of radiological studies 2/7/2013 63
  • 64. Electrophysical Technologies ā€¢ Various functional disturbances sometimes detectable only via analysis of electric potentials produced by some organs (tissues) as a manifestation of their function (ECG, EEG, EMG, ERG and others). ā€¢ Evaluation of these signals is performed either in the ā€œtime domainā€ (detection of particular specific peaks and wave complexes - ā€œgrapho-elementsā€ - and description of their time and amplitude characteristics) or in the ā€œfrequency domainā€ (frequency spectrum characteristics). ā€¢ At present this is done almost exclusively with the use of computers. 2/7/2013 64
  • 65. Electrophysiology contdā€¦ Electroencephalography : ā€¢ This is the neurophysiologic measurement of the electrical activity of the brain by recording from electrodes placed on the scalp or in special cases, subdurally or in the cerebral cortex. It represent an electrical signal (postsynaptic potentials) from a large number of neurons. ā€¢ Each electrode is connected to an input of a differential amplifier, which amplifies the voltage between them. The resulting voltage signal is filtered which is shown on paper or displayed on a computer screen. Uses in ā€“ Epilepsy and syncope ā€“ Sleep disorders ā€“ Coma and brain death 2/7/2013 65
  • 66. Electrophysiology contdā€¦ Electrocardiography : An electrocardiogram records the electrical activity of the heart over time. ā€¢ Electrodes on different sides of the heart measure the activity of different parts of the heart muscle. ā€¢ An ECG displays the overall rhythm of the heart and weaknesses in different parts of the heart muscle. ā€¢ Applications in public health : It is the best way to measure and diagnose abnormal rhythms of the heart in ā€“ Damage to the conductive tissue carrying electrical signals, ā€“ High or low potassium. ā€“ Myocardial infarction (MI) 2/7/2013 66
  • 67. Electrophysiology contdā€¦ Electromyography : ā€¢ Electromyography (EMG) is a technique for evaluating and recording the activation signal of muscles. ā€¢ An electromyograph detects the electrical potential generated by muscle cells when these cells contract and also when the cells are at rest. ā€¢ EMG is used to diagnose two general categories of disease : neuropathies and myopathies. 2/7/2013 67
  • 68. ENDOSCopy ā€¢ Different types of endoscopies are performed for diagnostic purposes of carcinoma and other pathologies. These may be rigid or fibreoptic. ā€¢ Direct:- ā€“ Laryngoscopy ā€“ Broncoscopy ā€“ Colonoscopy ā€“ Gastroscopy ā€“ Nasopharyngoscopy ā€“ Sigmoidoscopy ā€“ Hysteroscopy ā€¢ Indirect:- ā€“ Laryngoscopy 2/7/2013 68
  • 69. References ā€¢ Rajvir Balwar, Text book Public Health and Community Medicine, 1st Edition, AFMC Pune in collabration with WHO, Dept Of Commnity Medicine, 2008. ā€¢ Indian Public Health Standards (IPHS) Guidelines for Sub- District/Sub-Divisional Hospitals (31 to 100 Bedded) Revised 2012 , DGHS, MOHFW, Govt Of India, June 2012. ā€¢ Indian Public Health Standards (IPHS) Guidelines for District Hospitals (101 to 500 Bedded) Revised 2012, DGHS, MOHFW, Govt Of India, June 2012. ā€¢ http://www.biomerieux.com/en/advanced-technologies ā€¢ World Health Organization, Laboratory Support to Emerging Diseases. World Health Organization Regional Office for South-East Asia New Delhi, 2008. ā€¢ www.whoindia.org/LinkFiles/Public_Health_Laboratory_Networkin g_SEARO_recommendations _on_networking_of_PHL 2/7/2013 69
  • 70. ā€¢ The Diagnostics Innovation Map: Medical Diagnostics for the Unmet Needs of the Developing World, WHO BIO Ventures for Global Health,2010. ā€¢ Estimating the Global Health Impact of Improved Diagnostic Tools for the Developing World. Santa Monica, Rand Institute,2007. ā€¢ Sen J, Belli A "S100B in neuropathologic states: the CRP of the brain?". J. Neurosci. Res.2007; 85 (7): 1373ā€“80. ā€¢ http://mohfw.nic.in/NRHM/iphs.htm ā€¢ http://testequipment.globalspec.com/LearnMore/Labware_Scientif ic_Instruments/Analytical_Instruments/Chromatographs/HPLC ā€¢ http://www.google.co.in/imgres?um/diagnostic%technologies/php. 2/7/2013 70
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