PUS, GENITAL AND BODY FLUID BENCH
PUS SPECIMEN
Method
- This procedure consists of direct smear examination and culture of the affected sire. Smear prepared from swab or exudative material and subsequently Gram stained may show the presence of inflammatory cells as well as characteristic morphology that may lead to an initial diagnosis.
- Agent of primary infection are recovered in routine culture using primary non-selective media such as Blood Agar and selective media such as MacConkey Agar. This procedure also addresses isolation and identification of the striae anaerobic from aspirated pus collection.
- Biochemical identification according to species that been suspected
- Antimicrobial susceptibility testing.
Common Pathogen
Staphylococcus aureus, Streptococcus pyogenes, Enterococci, Anaerobic
Streptococcus, Clostridium tetani, Escherichia coli,Klebsiella spp. Pseudomonas
aeruginosa, Proeus spp, Pasteurella spp.
EAR AND EYE SWAB SPECIMEN
Common Pathogen
In Eye Infection
Gram Positive Organism
|
Gram Negative Organism
|
Staphylococcus aureus
Streptoccus pneumoniae
Streptococcsagalactie(Group B)-Eye infection of newborn
Streptococcus pyogenes and other beta-haemolytic
Streptococci
|
Neisseria gonorrhoeae-eye infection of newborn
H. influenzae
Pseudomonas
aeruginosa
Moraxella
lacunata
Enterobactericeae
|
In Ear Infection
External Ear Infections
|
Middle Ear Infections
|
Inner Ear Infections
|
Staphylococcus aureus
Streptococcus pyogenes
P. aeruginosa
|
S. aureus
S.pneumoniae
S. pyogenes and other
beta-haemolytic streptococcus
Klebsiella spp
P.aeruginosa
H.Influenzae
|
Bacteriodes.spp
Fungi
|
Principle Of Test
- This procedure consist of direct snear examination and culture of the affected sire. Smear prepared from swab or exudative material and subsequently Gram stained may show the presence of inflammatory cells as well as characteristic morphology that may lead to an initial diagnosis.
- Agent of primary infection are recovered in routine culture using primary non-selective media such as Blood Agar, enrich media such as Chocolate Agar and selective media such as MacConkey Agar and Sabouraud Dextrose Agar with Chloramphenicol + Gentamicin.
- Biochemical identification according to species that been suspected
- Antimicrobial susceptibility testing.
GENITALIA SPECIMEN
Purpose: Culture of
genitalia specimen. Common pathogen are Neisseria gonnorhea, Candida Albicans,
Strep Group B, Trichomonas vaginalis and urethritis.
Method
Direct smear with Gram Staining-Culture
on media plate - Blood agar, MacConkey Agar, Sabouraud Dextrose Agar +
Chloramphenicol and Gentamicin and Chocolate agar (incubate overnight)
Biochemical identification
Antimicrobial susceptibility
testing
BODY FLUID SPECIMEN
Principle of the method. The principle
of the test is by macro and microscopically examination, cell count, isolation,
identification and susceptibility testing of organism.
Common Pathogen
Gram Positive Organism
|
Gram Negative Organism
|
Staphylococcus aureus
Staphylococcus epidermidis
Streptococcus pneumoniae
Streptococcus pyogenes
Anaerobic Streptococci
|
Escherichia coli
Klebsiella spp. Enterobacter
spp
Pseudomonas spp
Pseudomonas aeruginosa
Other Enterobactericeae
|
Others;
Yeast
Fungi
|
Method
- Describe appearance of the sample; clear, turbid, xanthochromic or blood stain
- Perform cell count for number of white cell in sample
- Centrifuge sample at 3000rpm for 20 minutes.
- Decant supernatant and leave 0.5 ml and vortex for 5 seconds
- Inoculate one drop of specimen onto Blood agar, MacConkey agar and Anaerobic blood agar with Metronidazole disc and 1 drop on clear slide for gram staining.
- Incubate 18 to 24 hour in incubator at 35 to 370C for blood agar and MacConkey agar and incubate 48 hour for anaerobic agar in anaerobic jar at 35 to 370C.
- Read identification and perform biochemical identification for confirmation and antimibrobial susceptibility test.
INOCULATION TECHNIQUE
Inoculation been done in media
plate agar for identification of bacterial colony morphology purpose.
ANTIMICROBIAL SUSCEPTIBILITY TESTING
Purpose: Identification of
bacterial sensitivity to certain antibiotic. Antimicrobial susceptibility test is
performed on bacteria isolated from clinical specimen of isolate is determined
to be the pathogens. Susceptibility testing is required to assist the clinician
in the choice of agents for therapeutic or prophylactic use.
Reagent And Material Used
- Mueller Hinton broth
- Mueller Hinton agar
- Mueller Hinton blood agar for fastidious organism
- Antibiotic Disc
Method
- Using sterile swab, take 2-3 colony with same morphology and transfer into Mueller Hinton broth and mix
- With same sterile swab, place Mueller Hinton Broth onto whole Mueller Hinton agar accordingly.
- Using a needle, place antibiotic on the agar evenly according to type of the bacteria.
Result And Interpretation
- Sensitive-Zone of inhibition diameter> 12mm
- Partial sensitive-Zone of inhibition diameter
- Resistance-Zone of inhibition diameter
STERILITY CULTURE
Purpose: to monitor the sterility
of pharmaceutical product prepared by the hospital’s pharmacy department.
Principle: Contaminated
pharmaceutical preparation may cause serious infections if used on patient.
Therefore, samples of each batch of pharmaceutical product prepared in the
hospital’s pharmacy department are sent to microbiology laboratory for testing.
Samples are diluted and cultured to check for sterility
CULTURE FOR ENVIRONMENT
Purpose: To detect the
presence of microorganism in the environment and occurrence of nosocomial
disease develop in hospital
Principle: Environment
swab are taken from operation theatre and intensive cae units to ascertain the
organism that cause nosocomial infection to patients.
CULTURE FOR IN USE TEST
Purpose: To test the
efficiency and usage of disinfectant in the laboratory and hospital
Principle: In used
disinfectant solution from laboratory and hospital are tested for efficiency
process of disinfection that is being practiced.
CULTURE FOR IN USE TEST
Purpose: To test the
efficiency and usage of disinfectant in the laboratory and hospital
Principle: In used
disinfectant solution from laboratory and hospital are tested for efficiency
process of disinfection that is being practiced
STAINING
Staining method that always been
used in this lab are
Gram Stain
Purpose: To identify gram
positive organism from gram negative organism. It is a primary identification
of bacterial presence in patient sample before being further tested.
Reagent Needed
Crystal violet (primary staining)
Iodine crystal (megukuhkan
pewarnaan)
Aseton (decolorizer)
Safranin (counter staining)
Result
Gram Positive Organism-Blue/Purple
Gram Negative Organism-Red
Acid Fast Stain
Purpose: To differentiate
acid fast bacillus fron non-acid fast bacillus. It is useful for primary
identification of mycobacterium tuberculosis that can cause tuberculosis
Reagent Needed
a)Carbol Fuchsin (need heating
to allow staining to enter microorganism)
Alcohol (decolorizer)
Methylene blue (counterstain)
Result
Pink-acid fast bacillus
Blue-nonacid fast bacillus
b)India Ink Stain
Purpose: It is used to
enhance th mcroscopic detection of Cryptococcus
spp. in wet preparation. It is a negative stain resulting in a dark
backgroung and unstained organisms. It is important for CSF specimen because Cryptococcus neoformans is the most
cause of fungal CNS infections
Method
1 drop of CSF sample dropped on
clear glass slide.
Add India ink reagent on to the
CSF sample.
Cover with coverslit.
Check under microscope
Results/Interpretation
Positive - Cryptococcus neoformans
Negative- Candida albicans
c)Lactophenol Cotton Blue
Stain
Purpose: Lactophenol
Cotton Blue Stain Dropper are intended for use as in examining of fungi. Upon
the addition of lactophenol cotton blue, fungi stain blue allowing easier
visualization and examination
Method
Prepare wet mount with the stain
Results/Interpretation
Yeast cells, mycelia and fruiting
structures stain a delicate blue color, while the background appears as faint,
pale blue.
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