This document provides information on the history, epidemiology, microbiology, pathogenesis, diagnosis and clinical features of tuberculosis. Some key points:
- Tuberculosis is an ancient disease that has affected humans for thousands of years. Robert Koch discovered the causative bacteria, Mycobacterium tuberculosis, in 1882.
- In 2020, there were an estimated 10 million new TB cases and 1.5 million TB deaths worldwide, making it one of the top 10 causes of death. India has the highest burden of cases.
- M. tuberculosis is an aerobic bacterium with a complex cell wall structure that allows it to be acid-fast staining. It typically causes a chronic pulmonary infection but can spread to other
Tuberculosis is a chronic, wasting, communicable disease, which made a huge comeback with the HIV pandemic, making it an opportunistic infection, and and an AID-defining infection. This presentation explores the different types of tuberculosis in terms of their locations (pulmonary and extra-pulmonary) as well as in terms of their drug susceptibility. It also addresses the approach to the management of each one of these.
Tuberculosis is a chronic, wasting, communicable disease, which made a huge comeback with the HIV pandemic, making it an opportunistic infection, and and an AID-defining infection. This presentation explores the different types of tuberculosis in terms of their locations (pulmonary and extra-pulmonary) as well as in terms of their drug susceptibility. It also addresses the approach to the management of each one of these.
Pediatrics notes about "Tuberculosis". These notes were published in 2018.
You can download them also from
- Telegram: https://t.me/pediatric_notes_2018
- Mediafire: http://www.mediafire.com/folder/u5u60m184t9z7/Pediatric_Notes_2018
Tuberculosis is a communicable chronic granulomatous disease caused by Mycobacterium tuberculosis , where the center of the granuloma is Caseous necrosis
It usually involves the lungs but may affect any organ or tissue in the body
Airborne spread of droplet nuclei
More than 5.7 million new cases of TB (all forms, both pulmonary and extra-pulmonary) were reported to the World Health Organization (WHO) in 2013; 95% of cases were reported from developing countries
Latest figures from 20151 indicate an estimated 10.4 million people had TB, and 1.8 million people died (1.4 million HIV negative and 400 000 HIV positive).
Of further concern is that 480 000 cases of multidrug-resistant (MDR) TBa and a further 100 000 that were estimated to be rifampicin-resistant (RR) TB have occurred in the same period.
A presentation about Tuberculosis . This presentation composed of the definition, causes, pathophysiology, clinical feature, diagnosis, treatment, prognosis and prevention of Tuberculosis.
Tuberculosis (TB) is an infectious disease usually caused by Mycobacterium tuberculosis (MTB) bacteria. Tuberculosis generally affects the lungs, but can also affect other parts of the body. Most infections show no symptoms, in which case it is known as latent tuberculosis.
TB is spread from person to person through the air. When people with lung TB cough, sneeze or spit, they propel the TB germs into the air. A person needs to inhale only a few of these germs to become infected.
Pediatrics notes about "Tuberculosis". These notes were published in 2018.
You can download them also from
- Telegram: https://t.me/pediatric_notes_2018
- Mediafire: http://www.mediafire.com/folder/u5u60m184t9z7/Pediatric_Notes_2018
Tuberculosis is a communicable chronic granulomatous disease caused by Mycobacterium tuberculosis , where the center of the granuloma is Caseous necrosis
It usually involves the lungs but may affect any organ or tissue in the body
Airborne spread of droplet nuclei
More than 5.7 million new cases of TB (all forms, both pulmonary and extra-pulmonary) were reported to the World Health Organization (WHO) in 2013; 95% of cases were reported from developing countries
Latest figures from 20151 indicate an estimated 10.4 million people had TB, and 1.8 million people died (1.4 million HIV negative and 400 000 HIV positive).
Of further concern is that 480 000 cases of multidrug-resistant (MDR) TBa and a further 100 000 that were estimated to be rifampicin-resistant (RR) TB have occurred in the same period.
A presentation about Tuberculosis . This presentation composed of the definition, causes, pathophysiology, clinical feature, diagnosis, treatment, prognosis and prevention of Tuberculosis.
Tuberculosis (TB) is an infectious disease usually caused by Mycobacterium tuberculosis (MTB) bacteria. Tuberculosis generally affects the lungs, but can also affect other parts of the body. Most infections show no symptoms, in which case it is known as latent tuberculosis.
TB is spread from person to person through the air. When people with lung TB cough, sneeze or spit, they propel the TB germs into the air. A person needs to inhale only a few of these germs to become infected.
MANAGEMENT OF ATRIOVENTRICULAR CONDUCTION BLOCK.pdfJim Jacob Roy
Cardiac conduction defects can occur due to various causes.
Atrioventricular conduction blocks ( AV blocks ) are classified into 3 types.
This document describes the acute management of AV block.
These simplified slides by Dr. Sidra Arshad present an overview of the non-respiratory functions of the respiratory tract.
Learning objectives:
1. Enlist the non-respiratory functions of the respiratory tract
2. Briefly explain how these functions are carried out
3. Discuss the significance of dead space
4. Differentiate between minute ventilation and alveolar ventilation
5. Describe the cough and sneeze reflexes
Study Resources:
1. Chapter 39, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 34, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 17, Human Physiology by Lauralee Sherwood, 9th edition
4. Non-respiratory functions of the lungs https://academic.oup.com/bjaed/article/13/3/98/278874
Anti ulcer drugs and their Advance pharmacology ||
Anti-ulcer drugs are medications used to prevent and treat ulcers in the stomach and upper part of the small intestine (duodenal ulcers). These ulcers are often caused by an imbalance between stomach acid and the mucosal lining, which protects the stomach lining.
||Scope: Overview of various classes of anti-ulcer drugs, their mechanisms of action, indications, side effects, and clinical considerations.
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
- Video recording of this lecture in Arabic language: https://youtu.be/Ve4P0COk9OI
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
Factory Supply Best Quality Pmk Oil CAS 28578–16–7 PMK Powder in Stockrebeccabio
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Report Back from SGO 2024: What’s the Latest in Cervical Cancer?bkling
Are you curious about what’s new in cervical cancer research or unsure what the findings mean? Join Dr. Emily Ko, a gynecologic oncologist at Penn Medicine, to learn about the latest updates from the Society of Gynecologic Oncology (SGO) 2024 Annual Meeting on Women’s Cancer. Dr. Ko will discuss what the research presented at the conference means for you and answer your questions about the new developments.
Explore natural remedies for syphilis treatment in Singapore. Discover alternative therapies, herbal remedies, and lifestyle changes that may complement conventional treatments. Learn about holistic approaches to managing syphilis symptoms and supporting overall health.
Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
micro teaching on communication m.sc nursing.pdfAnurag Sharma
Microteaching is a unique model of practice teaching. It is a viable instrument for the. desired change in the teaching behavior or the behavior potential which, in specified types of real. classroom situations, tends to facilitate the achievement of specified types of objectives.
New Directions in Targeted Therapeutic Approaches for Older Adults With Mantl...i3 Health
i3 Health is pleased to make the speaker slides from this activity available for use as a non-accredited self-study or teaching resource.
This slide deck presented by Dr. Kami Maddocks, Professor-Clinical in the Division of Hematology and
Associate Division Director for Ambulatory Operations
The Ohio State University Comprehensive Cancer Center, will provide insight into new directions in targeted therapeutic approaches for older adults with mantle cell lymphoma.
STATEMENT OF NEED
Mantle cell lymphoma (MCL) is a rare, aggressive B-cell non-Hodgkin lymphoma (NHL) accounting for 5% to 7% of all lymphomas. Its prognosis ranges from indolent disease that does not require treatment for years to very aggressive disease, which is associated with poor survival (Silkenstedt et al, 2021). Typically, MCL is diagnosed at advanced stage and in older patients who cannot tolerate intensive therapy (NCCN, 2022). Although recent advances have slightly increased remission rates, recurrence and relapse remain very common, leading to a median overall survival between 3 and 6 years (LLS, 2021). Though there are several effective options, progress is still needed towards establishing an accepted frontline approach for MCL (Castellino et al, 2022). Treatment selection and management of MCL are complicated by the heterogeneity of prognosis, advanced age and comorbidities of patients, and lack of an established standard approach for treatment, making it vital that clinicians be familiar with the latest research and advances in this area. In this activity chaired by Michael Wang, MD, Professor in the Department of Lymphoma & Myeloma at MD Anderson Cancer Center, expert faculty will discuss prognostic factors informing treatment, the promising results of recent trials in new therapeutic approaches, and the implications of treatment resistance in therapeutic selection for MCL.
Target Audience
Hematology/oncology fellows, attending faculty, and other health care professionals involved in the treatment of patients with mantle cell lymphoma (MCL).
Learning Objectives
1.) Identify clinical and biological prognostic factors that can guide treatment decision making for older adults with MCL
2.) Evaluate emerging data on targeted therapeutic approaches for treatment-naive and relapsed/refractory MCL and their applicability to older adults
3.) Assess mechanisms of resistance to targeted therapies for MCL and their implications for treatment selection
2. History of Tuberculosis
Tuberculosis- an ancient disease
“Captain among these men of death”
Signs of Skeletal TB have been found in
remains from Europe from Neolithic times
(8000 BC), ancient Egypt (1000 BC)
TB was recognized as a contagious disease
by the time of Hippocrates( 400 BC) –
termed as “phthisis”
German physician Robert Koch discovered
and isolated Mycobacterium tuberculosis in
1882
3. Key Facts of the Disease
• A total of 1.5 million people died from TB in 2020
• In 2020, there were an estimated 10 million new incident cases of TB
worldwide (WHO)
• Men 56 Lakh
• Women 33 Lakh
• Children 11 Lakh
• The five countries with the highest number of cases were India,
China, South Africa, Indonesia, and Pakistan
• An estimated 6.6 crore lives were saved through TB diagnosis and
treatment between 2000 and 2020
4. India has the highest burden of TB in the world
•Incidence of 27 lakh new cases in 2019
•Incidence rate of 199 cases per lakh
population
5. General characteristics
• Tubercle bacilli can remain viable for many years in the tissues of
healthy persons
• When they produce disease, it runs a chronic and protracted course
that gives ample time for transmission to susceptible hosts
• The infection can produce disease in a human being after decades of
dormancy
• It can produce an epidemic, when introduced into a population of
which only a small portion is immunologically protected by already
having been infected
6. M. tuberculosis complex (MTBC)
• The most common causative agent of TB in humans, Mycobacterium
tuberculosis, is a member of the M. tuberculosis complex (MTBC) which includes
six other closely related species:
• M. bovis - causes disease in cattle and spreads to humans through animal
contact and consumption of unpasteurized milk
• M. bovis BCG variant
• M. africanum - causes human TB in West Africa, up to 50% cases
• M. microti
• M. pinnipedii
• M. caprae
• M. canettii
All MTBC members are obligate pathogens and cause TB
7. M. tuberculosis
• Aerobic
• non–spore-forming
• nonmotile bacillus
• with a high cell wall content of high-molecular-weight lipids.
• Growth is slow - generation time being 15 to 20 hours,
compared with much less than 1 hour for most common
bacterial pathogens
• Visible growth takes from 3 to 8 weeks on solid media
8. Live culture colonies
in L J Medium
• The colonies of Mycobacterium on solid
culture media can be of various shades from
cream to yellow and deep orange color
• M. tuberculosis produces cream-colored buff,
rough colonies after 2–3 weeks inoculation on
Löwenstein–Jensen medium
9. Live culture colonies in LJ Medium
• Smooth colonies of
Mycobacterium
grown on
Löwenstein–Jensen
medium
• Rough colonies of
Mycobacterium grown
on Löwenstein–Jensen
medium
11. Acid-fast staining
• The term acid-fast bacilli (AFB) is practically synonymous with
mycobacteria, although Nocardia and some other organisms
are variably acid-fast
• The cell walls of mycobacteria, because of their high lipid
content, have the unique capability of binding fuchsin dye so
that it is not removed (destained) by acid–alcohol
12. Pulmonary Tuberculosis
• The lung is the most commonly affected organ in TB infection in the
immunocompetent host
• Lung involvement in subjects with active tuberculosis of 80%
• The lung is the portal of entry in the majority of cases of tuberculosis
• The first contact with the organism results in few or no clinical
symptoms or signs
• Ordinarily, the tubercle bacillus sets up a localized infection in the
periphery of the lung, where it has been deposited by inhalation
• Body defenses appear to have little effect on the organism until the
time of development of tuberculin hypersensitivity (4 to 6 weeks)
13. Pulmonary Tuberculosis
• Pulmonary tuberculosis frequently develops slowly, without a definite date
of onset
• Symptoms may be divided into two categories –
• Constitutional – fever, weight loss, fatigue
• Pulmonary – cough, expectoration (sputum production), hemoptysis, chest pain
• Characteristically, the fever develops in the late afternoon and may not be
accompanied by pronounced symptoms.
• With defervescence, usually during sleep, sweating occurs—the classic
“night sweats.”
14.
15. Pulmonary Tuberculosis
• Apical-posterior localization with a tendency to cavitation and
progression is characteristic of pulmonary TB in adolescents and adults
• Apical localization of pulmonary TB (adults) –
• the hyperoxic environment of the apices and the aerobic nature of the
organism.
• Deficient lymphatic flow at the lung apices, especially the posterior apices,
where the pumping effect of respiratory motion is minimal.
• Deficient lymph traffic would favor retention of bacillary antigen and, when
hypersensitivity ensues, tissue necrosis.
• Elderly and progressive primary infection of childhood - often causes
nondescript lower lobe pneumonia
16. Extra Pulmonary Tuberculosis
Lymph node tuberculosis
Pleural effusion and empyema thoracis
Bone and joint tuberculosis
Genitourinary tuberculosis
Disseminated tuberculosis
CNS tuberculosis
Abdominal tuberculosis
Others
17. Tuberculous
Lymphadenitis
• Most common presentation of EPTB in both HIV
infected and non infected patients
• Accounts for nearly 35% of EPTB cases
• 50% cases involve peripheral lymph nodes
• Posterior cervical and supraclavicular sites are
commonly involved
• Cervical lymphadenopathy is common in HIV-negative
• Multifocal involvement common in HIV-positive
18. Tubercular Pleural Effusion
• Accounts for nearly 20% of EPTB cases
• Usually presents as an acute illness and symptom
duration ranges from a few days to few weeks
• A small sub pleural focus may rupture into the
pleural space
• Present with chest pain, breathlessness, cough
with expectoration, fever, and toxaemia
• Occasionally, tuberculosis empyema may present
as a chest wall mass or draining sinus tract
19. Skeletal TB
• Accounts for nearly 10% of EPTB
• Reactivation of hematogenous foci or
spread from adjacent paravertebral
lymph nodes
• Spine (40%), hips (13%), knee (10%)
commonly affected
• Lower thoracic and lumbar vertebrae
are commonly involved followed by
middle thoracic and cervical vertebrae
20. CNS TB & Tuberculosis meningitis
• Accounts for approx 1 - 5% of cases of EPTB
• Results from hematogenous spread of primary or pulmonary TB or
from rupture of a subependymal tubercle into subarachnoid space
• Pathological features of TBM:
• Inflammatory meningeal exudate
• ependymitis
• vasculitis
• encephalitis
• disturbance of CSF circulation and absorption
• With onset of meningitis - headache, neck rigidity and vomiting, fever
develops
21. CNS TB & Tuberculosis meningitis
• Focal neurological deficits and features of raised intracranial tension may
precede signs of meningeal irritation
• Focal or generalised seizures- 20 to 30 % of patients
• Cranial nerve palsies- 20 to 30 % of patients, 6th nerve involvement is
most common
• Hydrocephalus occurs in approximately two-third of cases
• Complete or partial loss of vision- a major complication of TBM
• Terminal illness is characterized by deep coma and decerebrate/
decorticate posturing
25. Gastrointestinal Tuberculosis
• 70% of patients with advanced pulmonary disease acquired gastrointestinal TB
from swallowing infectious secretions
• Abdominal TB can present in any of the sites:
Most commonly -
• Peritoneum
• intestinal tract
• lymph nodes
Also may involve -
• Stomach
• hepatobiliary tree
• Pancreas
• perianal area
26. Latent TB infection - Definition
• The Latent Tuberculosis Infection (LTBI) has been defined as infection
with M. tuberculosis within granuloma, that remains in non
replicating state but retains its ability to come out of latency and
cause active TB if and when a disruption of the immune response
occurs.
27. Difference Between Latent TB Infection and
TB Disease
Latent Tuberculosis Active Tuberculosis
• Have no symptoms
• Do not feel sick
• Cannot spread TB to others
• Usually have a positive skin and IGRA
test
• Chest x-ray and sputum test usually
normal
•Symptoms include
a) bad cough that lasts longer than 2 weeks
b) pain in the chest
c) coughing up blood or sputum
d) weakness or fatigue
e) weight loss
f) no appetite
g) chills, fever, sweating at night
• May spread TB to others
• Usually have a positive skin and IGRA
test
• May have abnormal chest x-ray, and/or
positive sputum smear or culture
28. Tests available for LTBI
There are currently two major classes of tests used to identify
patients with latent tuberculosis:
1) The tuberculin skin tests include:
• Mantoux test
2) The IFN-γ (interferon-gamma release assay - IGRA) tests include:
• QuantiFERON-TB Gold
• T-SPOT TB
29. Increased susceptibility to TB
A. Nonspecific decrease in resistance
• < 5 yrs of age and close contact of TB
• Senescence
• Malnutrition
• Postgastrectomy state
• Diabetes mellitus
• Renal failure
• Smoking, alcohol & drug abuse
B. Decrease in resistance due to hormonal
effects
• Pregnancy
• Therapy with adrenocortical steroids
C. Decrease in local resistance
• Silicosis
D. Decrease in specific immunity
• Lymphomas, Head & Neck cancers
• Immunosuppressive therapy (anti TNF⍺,
corticosteroids)
• Sarcoidosis
• HIV infection
• Transplantation
30. HIV-TB
• TB- leading cause of HIV/AIDS related mortality globally
• Risk of TB 16-27 times higher in PLHA, almost 60% of TB was not
diagnosed/treated
• Frequently disseminated disease, patient asymptomatic and TB goes
undetected- need for screening
• Low immunity- less cavitation/ sputum production
• Difficulty in procuring sputum / respiratory samples
• Technical expertise required in tests such as Xpert, LPA
32. Specimen collection for Diagnosis
• Mycobacteria can be recovered from a variety of clinical specimen -
Respiratory specimens (sputum, bronchial washes,
bronchoalveolar lavage fluid, and bronchial biopsies)
Urine, feces, blood, cerebrospinal fluid;
Tissue biopsies; and
Deep-needle aspirations of virtually any tissue or organ
• Specimens that may contain normal bacterial flora should be
processed as soon after collection as possible to minimize the degree
of overgrowth with specimen contaminants
33. Specimen - Sputum
• Sputum samples collected by expectoration or by ultrasonic
nebulization are best obtained shortly after the patient awakens in
the morning, when mycobacteria are at their highest concentration
• Irregular and intermittent release of mycobacteria into the bronchial
lumen from mucosal ulcers or loculated cavities often results in a
variable pattern of recovery from respiratory secretions
34. Lab methods
• Digestion & Decontamination procedures for sputum :
• The high concentration of lipids in the cell wall of most
mycobacteria makes them more resistant to killing by strong
acid and alkaline solutions than other bacteria that may be
present in the specimen.
• Consequently, specimens likely to contain a mixed bacterial
microbiota are treated with a decontaminating agent to reduce
undesirable bacterial overgrowth and to liquefy mucus
a) Petroff’s method (NaOH)
b) NALC – NaOH method (N acetyl L Cystine)
35. Direct Smear Microscopy
• Ziehl-Neelsen (ZN) staining / Acid Fast
• Kinyoun stain – cold
• Fluorescent staining (Rhodamine/ Auramine O)
–5 X 103 to 5 X 104 bacilli / ml of sputum is required
for detection by smear
–Culture detects 10 to 100 viable organism
37. • Solid media
a) Egg based - L J, Petragnani, Dorset
b) Agar based - media are transparent; colonies observed in 10 to 12 days
in contrast to 18 to 24 days with opaque egg based media.
eg. Middle brook 7H11, Middle brook 7H10
Culture media
40. MGIT - Principles
• MGIT tube contains Middlebrook 7H9 liquid media and an
oxygen-quenched fluorochrome embedded in silicone at
bottom of the tube
• Bacterial growth = free oxygen is utilized and replaced with
carbon dioxide
• With depletion of free oxygen, the fluorochrome is no longer
inhibited, resulting in fluorescence within the MGIT tube when
visualized under UV light
• The intensity of fluorescence is directly proportional to the
extent of oxygen depletion.
41. MGIT …contd
• An antimicrobial mixture called PANTA™ (Polymyxin B,
Amphotericin B, Nalidixic Acid, Trimethoprim, Azlocillin) is used
for reducing the contamination of other bacteria
• MGIT system has also been used to detect drug susceptibility
against Antitubercular drugs – particularly for SIRE
(Streptomycin, Isoniazid, Rifampicin and Ethambutol)
42. MPT 64 Antigen test
• MPT64 is one of the major MTB complex culture filtrate protein
• This lateral flow test has been reported to identify the M.
tuberculosis complex from the MOTT in the Liquid Culture using the
mouse monoclonal anti-MPT64 antibody
43. Lab Diagnosis of Pulmonary TB
• Sample collection
• Sample decontamination and liquefaction
• Microscopy – ZN Staining / Fluorescence staining
• NAAT – GeneXpert / TrueNAAT
• Culture – Solid medium – LJ Medium
Liquid Medium – Bactec MGIT 960
• Drug Susceptibility – Solid media / liquid media / NAAT
• NAAT DST – Line Probe Assay, GeneXpert
45. Gene Xpert
• Single use cartridge based nucleic acid amplification test
(CBNAAT)
• Semi-quantitative, hemi-nested, real-time PCR, targeting the
81 bp Rifampin Resistance Determining Region (RRDR) of MTB
rpoB gene (96% of RR cases)
• Contains internal controls and five partially overlapping
fluorescent probes A, B, C, D and E
46. Time-to-result: 1 h 45 min
GeneXpert
DNA molecules are mixed
with dry PCR reagents
Sample is
automatically
filtered & washed
Ultrasonic lysis of filter-
captured organisms to
release DNA
Semi-nested real-time
amplification & detection
in integrated reaction tube
4
5
7
Concentrates bacilli &
removes inhibitors
1
2
Sputum liquefaction &
inactivation with 2:1 SR
Transfer of 2 ml
after 15 min
3
End of hands on work
6
Printable test result
Time-to-result: 1 h 45 min
GeneXpert
DNA molecules are mixed
with dry PCR reagents
Sample is
automatically
filtered & washed
Ultrasonic lysis of filter-
captured organisms to
release DNA
Semi-nested real-time
amplification & detection
in integrated reaction tube
4
5
7
7
Concentrates bacilli &
removes inhibitors
1
2
2
Sputum liquefaction &
inactivation with 2:1 SR
Sputum liquefaction &
inactivation with 2:1 SR
Transfer of 2 ml
after 15 min
3
End of hands on work
6
Printable test result
Gene Xpert
48. Line Probe Assay
• Multiplexed DNA hybridization assay - can interrogate a
variety of amplified genetic targets in a single, relatively
simple test format
• Available test kits: MTBDRplus V1, MTBDRplus V2, Nipro
NTM+MTBDR, MTBDRs V1/V2
• WHO’s stand on LPA:
– 2008: Use of FL-LPA (Genotype MTBDRplus)
– 2016: Recommended the use of SL-LPA (Genotype
MTBDRs)
Editor's Notes
Mycolic acid-
highly impermeable to antimicrobials such as penicillin
Provides hydrophobic mesh for other deeper complex lipids
LAM
D arabinosyl end capped: ManLAM- pathogenic mycobacteria- TB, leprae, bovis
Phosphoinositol capped: PILAM- non pathogenic- smegmatis
No capping- AraLAM- chelonae/ rapid growers
Mannan core highly conserved
Endorsed by who since 2010
In 2015, FIND (the Foundation for Innovative New Diagnostics) evaluated the Nipro and
the GenoType MTBDRplus V2 LPAs and compared them with the GenoType MTBDRplus
V1. The study demonstrated equivalence between these three commercially available
LPAs for detecting TB and resistance to Rif and H