This document discusses various epidemiological marker typing methods used to differentiate bacterial strains, including serotyping, bacteriocin typing, and colicin typing. Serotyping is based on antigenic differences expressed on bacterial cell surfaces and has good reproducibility but poor discriminatory power. Bacteriocin typing examines bacteriocin production and susceptibility patterns to distinguish strains. It has fair reproducibility and discriminatory power but some strains are non-typeable. Colicin typing specifically examines colicin production in E. coli strains using a spot culture method with indicator strains. These typing methods can help epidemiological studies and hospital infection control.
The use of a machine designed to follow repeatedly and automatically a predetermined sequence of individual operations.
AUTOMATED WASHING
AUTOMATED MEDIA PREPARATORS
AUTOMATED COLLECTION AND
PROCESSING OF SAMPLES
CYTOSPIN
AUTOMATED GRAM STAINING
AUTOMATED STREAKING
SPIRAL PLATER
AUTOMATED ANTIBIOTIC -
SENSITIVITY SYSTEM
AUTOMATIC COLONY COUNTER
AUTOMATED URINE MICROSCOPY -
ANALYSER
The use of a machine designed to follow repeatedly and automatically a predetermined sequence of individual operations.
AUTOMATED WASHING
AUTOMATED MEDIA PREPARATORS
AUTOMATED COLLECTION AND
PROCESSING OF SAMPLES
CYTOSPIN
AUTOMATED GRAM STAINING
AUTOMATED STREAKING
SPIRAL PLATER
AUTOMATED ANTIBIOTIC -
SENSITIVITY SYSTEM
AUTOMATIC COLONY COUNTER
AUTOMATED URINE MICROSCOPY -
ANALYSER
This is prepared for my project and im sharing this for useful to others.This slides contain the processing of urine specimens in microbiology.im prepared on basis of our medical college method.sometimes the methods will vary with other hospitals
Automated system for bacterial identificationDEEKSHANT KUMAR
[DOWNLOAD IT OPEN IT WITH MICROSOFT POWERPOINT THEN YOU WILL BE ABLE TO UNDERSTAND THE TOPIC COVERED.]
1. WHOLE TEXT IS RELIABLE.
2. TEXT HAS BEEN TAKEN FROM STANDARD TEXT BOOK FOR MEDICAL MICROBIOLOGY.
3. SOME PICTURE HAS BEEN TAKEN FROM JOURNAL.
Microbiology of E coli giving basic of Escherichia coli, its morphology, cultural and biochemical characteristics, Antigenic character, pathogenesis, laboratory diagnosis, prevention and control
This is prepared for my project and im sharing this for useful to others.This slides contain the processing of urine specimens in microbiology.im prepared on basis of our medical college method.sometimes the methods will vary with other hospitals
Automated system for bacterial identificationDEEKSHANT KUMAR
[DOWNLOAD IT OPEN IT WITH MICROSOFT POWERPOINT THEN YOU WILL BE ABLE TO UNDERSTAND THE TOPIC COVERED.]
1. WHOLE TEXT IS RELIABLE.
2. TEXT HAS BEEN TAKEN FROM STANDARD TEXT BOOK FOR MEDICAL MICROBIOLOGY.
3. SOME PICTURE HAS BEEN TAKEN FROM JOURNAL.
Microbiology of E coli giving basic of Escherichia coli, its morphology, cultural and biochemical characteristics, Antigenic character, pathogenesis, laboratory diagnosis, prevention and control
Production and applications of monoclonal antibodiesKaayathri Devi
production and applications of monoclonal antibodies, monoclonal antibodies ,applications of monoclonal antibodies, production of monoclonal antibodies,
What are Antibody
Monoclonal Antibody (mAb)
Structure of mAb
Types of Monoclonal Antibody (mAb)
Preparation of Monoclonal Antibody
Hybridoma Technique, Phage display Technique
Application of Monoclonal Antibody
Advantage and Disadvantage of Monoclonal Antibody
BIOTECHNOLOGY IS
CHALLENGING SUBJECT TO TEACH AND UNDERSTAND ......
ITS A VERY INTERESTING TO LEARN ABOUT HYBRIDOMA TECHNOLOGY .. THEIR PRODUCTION AND
APPLICATION ALSO ....
How many patients does case series should have In comparison to case reports.pdfpubrica101
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https://pubrica.com/academy/case-study-or-series/how-many-patients-does-case-series-should-have-in-comparison-to-case-reports/
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Defecation
Normal defecation begins with movement in the left colon, moving stool toward the anus. When stool reaches the rectum, the distention causes relaxation of the internal sphincter and an awareness of the need to defecate. At the time of defecation, the external sphincter relaxes, and abdominal muscles contract, increasing intrarectal pressure and forcing the stool out
The Valsalva maneuver exerts pressure to expel faeces through a voluntary contraction of the abdominal muscles while maintaining forced expiration against a closed airway. Patients with cardiovascular disease, glaucoma, increased intracranial pressure, or a new surgical wound are at greater risk for cardiac dysrhythmias and elevated blood pressure with the Valsalva maneuver and need to avoid straining to pass the stool.
Normal defecation is painless, resulting in passage of soft, formed stool
CONSTIPATION
Constipation is a symptom, not a disease. Improper diet, reduced fluid intake, lack of exercise, and certain medications can cause constipation. For example, patients receiving opiates for pain after surgery often require a stool softener or laxative to prevent constipation. The signs of constipation include infrequent bowel movements (less than every 3 days), difficulty passing stools, excessive straining, inability to defecate at will, and hard feaces
IMPACTION
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DIARRHEA
Diarrhea is an increase in the number of stools and the passage of liquid, unformed feces. It is associated with disorders affecting digestion, absorption, and secretion in the GI tract. Intestinal contents pass through the small and large intestine too quickly to allow for the usual absorption of fluid and nutrients. Irritation within the colon results in increased mucus secretion. As a result, feces become watery, and the patient is unable to control the urge to defecate. Normally an anal bag is safe and effective in long-term treatment of patients with fecal incontinence at home, in hospice, or in the hospital. Fecal incontinence is expensive and a potentially dangerous condition in terms of contamination and risk of skin ulceration
HEMORRHOIDS
Hemorrhoids are dilated, engorged veins in the lining of the rectum. They are either external or internal.
FLATULENCE
As gas accumulates in the lumen of the intestines, the bowel wall stretches and distends (flatulence). It is a common cause of abdominal fullness, pain, and cramping. Normally intestinal gas escapes through the mouth (belching) or the anus (passing of flatus)
FECAL INCONTINENCE
Fecal incontinence is the inability to control passage of feces and gas from the anus. Incontinence harms a patient’s body image
PREPARATION AND GIVING OF LAXATIVESACCORDING TO POTTER AND PERRY,
An enema is the instillation of a solution into the rectum and sig
India Clinical Trials Market: Industry Size and Growth Trends [2030] Analyzed...Kumar Satyam
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There is a movement towards home-based care for the elderly, next generation scanning and MRI devices, wearable technology, artificial intelligence incorporation, and online connectivity. Experts also see a focus on predictive, preventive, personalized, participatory, and precision medicine, with rising levels of integration of home care and technological innovation.
The average cost of treatment has been rising across the board, creating additional financial burdens to governments, healthcare providers and insurance companies. According to MCG, cost-per-inpatient-stay in the United States alone rose on average annually by over 13% between 2014 to 2021, leading MedTech to focus research efforts on optimized medical equipment at lower price points, whilst emphasizing portability and ease of use. Namely, 46% of the 1,008 medical technology companies in the 2021 MedTech Innovator (“MTI”) database are focusing on prevention, wellness, detection, or diagnosis, signaling a clear push for preventive care to also tackle costs.
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Empowering ACOs: Leveraging Quality Management Tools for MIPS and BeyondHealth Catalyst
Join us as we delve into the crucial realm of quality reporting for MSSP (Medicare Shared Savings Program) Accountable Care Organizations (ACOs).
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2. Introduction
Important to analyse multiple isolates within a given
species to determine whether they represent a single
strain or multiple strains.
If a species of bacteria is isolated and cultivated in the
laboratory it is known as a strain.
A single isolate with distinctive characteristic[s] may also
represent a strain.
Members of the same species that have small differences
between them can be distinguished by additional
methods. These species is then subdivided into
subspecies, subgroups, biotypes, serotypes, variants etc.
The process of differentiating strains based on their
phenotypic and genotypic differences is known as
'typing'.
3. These typing methods are useful in hospital infection
control, epidemiological studies, and understanding the
pathogenesis of infection.
In hospital settings they may be used to:
determine whether a set of isolates obtained from one
patient represents a single infecting strain or multiple
contaminants.
determine whether a series of isolates obtained over
time represents relapse of an infection due to single
strain or separate episodes of disease due to different
strains.
4. Criteria for evaluating typing systems:
1. Typeability: It is the ability to obtain unambiguous
positive result for each isolate analysed. Non-
typeable isolates are those that give either a null or
an uninterpretable result.
2. Reproducibility: It is the ability of the technique to
yield the same result when same strain is tested
repeatedly.
3. Discriminatory Power: It is the ability to
differentiate among unrelated strains. Ideally, each
unrelated isolate is detected as unique.
4. Ease of performance: To be widely useful, a typing
method should be applicable to a broad range of
microorganisms as well as inexpensive and
technically easy.
5. Ease of interpretation: The typings tests must
produce reproducible and unambiguous results that
5. Types of typing methods:
1. Phenotypic techniques, those that detect
characteristics expressed by the microorganism and
2. Genotypic techniques, those that involve direct DNA-
based analysis of chromosomal or extrachromosomal
genetic elements.
6. Serotyping
Based on fact that strains of same species can differ in
the antigenic determinants expressed on the cell
surface.
Surface structures such as lipopolysaccharides,
membrane proteins, capsular polysaccharides, flagella
and fimbriae exhibit antigenic variations and antibodies
raised against them can be used to group isolates into
defined serotypes.
Strains differentiated by antigenic differences are known
as 'serotypes'. Serotyping is used in several gram
negative and gram positive bacteria.
Some species are characterized by numerous antigenic
types and serotyping for these species is highly
discriminatory,
7. Serotyping is performed using several serologic tests
such as bacterial agglutination, latex agglutination,
co-agglutination, fluorescent and enzyme labelling
assays.
The following are the antigens used for developing
antibodies for serotyping.
"O" antigen- cell wall lipopolysaccharide antigen
Vi antigen
"H" antigen- protein antigen of flagella
"K" antigen- capsular polysaccharide antigen
8. For example,
Salmonella enterica - more than 2300 serotype based
on "O" antigen, Hantigen and Vi antigen (in Kauffman-
White scheme)
E. coli vary in the antigenic structure of certain parts of
the LPS portion of the cell wall, the O antigen. The
composition of the flagella, the H antigen, can also
vary..The strain designation of E. coli O157:H7 refers to the
structure of LPS and its flagella
Campylobacter jejuni- 55"O" antigens, 56"H" antigens,
80 "K" antigens
Klebsiella pneumoniae- 75 "K" antigens
Yersinia enterocolitica- 34 "O" antigens
Citrobacter freundii- 32 "O" antigens, 75 "H" antigens
Capsules, M and T proteins- Streptococcus pyogenes
Toxins- Clostridium spp.
9. Advantages:
Most strains are typeable. They have good
reproducibility and ease of interpretation though some
have ease of performance.
Disadvantages:
Some autoagglutinable (rough) strains are untypeable.
Some methods of serotyping are technically demanding.
There is dependency on good quality reagent from
commercial sources.
In-house preparation of reagents is difficult process.
Serotyping has poor discriminatory power due to large
number of serotypes, crossreaction of antigens and
untypeable nature of some strains.
11. BACTERIOCIN
11
Produced by many bacteria, protein in nature and
have bactericidal activity.
Have killing action on strains of same or closely
related species.
First reported by Gratia in 1925, Escherichia coli
producing a substances which is active against other
strains of the same species.
Colicin – Gratia and Frederique in 1946.
Bacteriocin- Jacob and Woolman in 1953.
12. contd…
12
Are named on the basis of their bacterial species of
origin . Some of them are
i) Colicins are bacteriocins of E.coli,
ii) Aeruginocin of P. aeruginosa,
iii) Diphthericins - C. diphtheriae
iv) Cloacin of Enterobacter cloaceae,
v) Pesticin of Y. pestis,
vi) Monocin of Listeria monocytogenes,
vii) Cerecin of Bacillus cereus
viii) Staphylococcin of Staphylococcus aureus,
ix) Warnerin of et S. warneri
13. contd…
13
Target cells have specific receptor for
attachment of bacteriocin( same as
bacterophage)
Bacteria producing bacteriocin also carry
gene for immunity to them on chromosome or
plasmid.
Many have narrow inhibitory spectrum of
activity ( but some have activity on broad
class of bacteria)
Are plasmid or chromosomal mediated.
14. Importance
14
Bacteriocins produced by non-pathogenic
bacteria kills other pathogenic
bacteria(Normal flora vs. Pathogens).
Bacteriocins have also been suggested for
certain cancer treatment.
Used for food presrvation in food industry (
eg..nisin produced by Lactococcus lactis is
active against many food spoiling bacteria)
Used for bacteriocin typing of clinical strains
to aid in their identification and
characterization.
15. Bacteriocin typing
15
A strain may be typed by:
1) Activity of their bacteriocin against a set of indicator
strains of same or closely related species, or
2) By strain against a set of bacteriocin which are
applied to it.
If the isolates are same strains, their bacteriocin
production and susceptibility patterns will be
identical.
The bacteriocin typing technique has been widely
used to answer the epidemiological questions in
the nosocomial infections.
16. Stab and overlay method
(Fredricq)
The test strains are stabbed in pre-poured BHI agar
or nutrient agar plates.
Incubated at 37ᴼC for 24 hours.
The plates are exposed to chloroform to kill the
producing strain. For this, a chloroform soaked filter
paper is placed at the lid of the plate and allowed to
stand for 10-15 minutes.
3 ml of BHI soft agar or nutrient agar containing 0.1
ml of standardized inoculum (2x108 cfu/ml) of
sensitive (indicator strain) culture is poured over the
plates and incubated at 37ᴼC for overnight.
After overnight incubation, zones of inhibition (ZOI)
around producer colonies are measured and
17. Cross streak method
The test strains are inoculated across the surface of
BHI agar plates.
After overnight incubation at 37ᴼC and exposing to
chloroform, the test (producing) strains are
scrapped-off from the plate.
The sensitive strains (Indicator strains) are cross-
streaked at right angle to the test strains.
Incubated at 37ᴼC for 24 hours.
Observed for inhibition of growth at each side of the
producing strain.
18. Preparation of cell free neutralized supernatant
(bacteriocin)
Producer strain is grown overnight at 37ᵒC in trypticase
soya broth.
0.25 ml of chloroform (for each 5ml of broth) is added to
kill the organisms.
1ml of mitomycin C can be added to induce the
production of bacteriocins.
Supernatant fluid is obtained by centrifuging at 10,000g
for 10 minutes.
pH of supernatant is adjusted to 7.0 with sterile 1N
NaOH.
19. Agar well diffusion method
Pre-poured BHI agar plates
are overlaid with 3ml of BHI
soft agar containing 0.1 ml
(2x108 CFU/ml) of the
sensitive (indicator) culture.
Wells (5 mm in diameter) are
cut into these agar plates.
100μl of the culture
supernatants (Bacteriocin) are
placed into each well and kept
at 4ᵒC for 10-12 hours to allow
the bacteriocin to diffuse into
the agar.
The plates are then incubated
at 37ᵒC for 24 hours.
20. Colicins
Colicins are bacterial macromolecules which have
narrow spectrum of activity and they are produced
by some strains of E. coli and related
Enterobacteriaceae.
They are specified by col plasmid.
They are synthesized in the cytoplasm of
colicinogenic cells and released into the extracellular
medium.
21. Colicin typing (Djonne)
Spot cultures are grown on plates of blood agar
base at 37ᵒC for 48 hours and killed by exposure to
chloroform vapour.
Leave at room temperature (RT) for 30 minutes to
remove residual chloroform.
One colony of universal indicator strain a1, grown
on blood agar, is mixed with 5 ml of 0.9% NaCl and
then overlaid on sterilized (with chloroform) plates.
After leaving for 1-2 hours at RT, they are incubated
for 24 hrs at 37ᵒC.
If the inhibition zone is >1mm around the spot
culture, colicin production is considered to be
positive.
22. Several biological criteria have been assessed
for typing Pseudomonas aeruginosa, including
- pigmentation,
- antibiograms and
- phage sensitivity.
However the two most reliable and
generally accepted methods are serotyping
and pyocin typing.
Pyocin typing
23. 23
Gillies and Govan(1966)- cross streaking
method
The test strains are inoculated across the
surface of BHI agar plates.
After overnight incubation at 37ᴼC , culture is
exposed to chloroform to kill the test strains and
then the test (producing) strains are scrapped-
off from the plate by using slide.
The sensitive strains (Indicator strains) are
cross-streaked at right angle to the test strains.
Incubated at 37ᴼC for 24 hours.
Observed for inhibition of growth at each side of
the producing strain.
24. Contd…
24
In pyocin typing technique with cross streaking
method, 105 main types and 25 subtypes can be
identified on the basis of pyocin production by
test strains using 13 indicators(1 to 8 and A to E).
Disadvantages of streak method
To remove test strains growth before application
of indicator strain.
Not reliable for mucoid colony of P. aeruginosa.
48 hour period is needed to obtain result.
25. Klebocin typing
Spot inoculation method as described by Chugh et al(1980)
using 6 producer strains 153,154,155,156, US-5 and US-6.
Each producer strain is cultured in a broth.
To each broth culture(0.7ml) of producer strains, 3.3 ml of
trypticase soya broth and 1ml of mitomycin C(0.5µgm/ml) is
added and incubated at 37C for 5 hours with intermittent
shaking.
The cells are killed by addition of 0.25 ml of chloroform.
The suspension is centrifuged at 3000rpm for 10 minutes in
cold centrifuge.
Supernatent containing respective klebocin is aliquoted in
screw cap vials and stored at -20C.
26. The test culture is swabbed over a nutrient agar plate.
12 µl of each klebocin is spot inoculated on marked
sectors with a micropipette.
A positive control plate streaked with strain WC-50 is
also put up with every set of test.
The negative and positive reactions to 6 klebocins are
recorded.
27. Advantages:
This technique has fair amount of reproducibility,
discriminatory power and ease of interpretation.
Disadvantages:
This technique is available only at reference centres.
Even for the experienced worker, the technique is
demanding
Many strains are non-typeable.