MICROBIOLOGY

L A B I N K . F. H
 Microbiology lab in
general

TOPICS
Microbiology lab in
K.F.H
Cases in K.F.H
 What is Microbiology Lab ?
It is about promoting the growth of bacteria to discover the type of bacteria that causes the disease and giving the
patient the appropriate antibiotic that kills the bacteria.

1- stimulate growth by 2- Genre discovery by 3- appropriate antibiotic by

Types of samples :
Any sample you can think of Whether it is from the human body, any body fluid such as urine, CSF, etc., or other
factors such as food
Microbiology lab in K.F.H
Microbiology lab in King Fahd Hospital is divided into two main sections:
❑ Parasitology section
❑ Bacterial culture section

➢ Parasitology section
In this section, two main samples are urine and stool
Urine is detected by urinalysis. This analysis is divided into two main parts:
biochemical test and microscopic examination.

1. Biochemical test :
Biochemical test is helpful in detecting UTI, It look for the presence of bacterial
enzymes or PMN enzymes.
Protein indication of bacteria or kidney problems
Nitrite indication of GNR
Leukocyte esterase indication of inflammation
 2. Microscopic examination
We are looking for:
• Bacteria
• WBC(White Blood Cells)
• RBC(Red Blood Cells)
• Epithelial Cells
• Yeast
• Casts (Hyaline, Waxy, Cellular and Granular)
• Crystals(Cystine , Triple phosphate& Calcium Oxalate)

Some interpretations:
• Large number of epithelial cells indicates UTIs or
vaginal contamination of the specimen
• Waxy casts indicates renal failure
• Red cells in casts indicates haemorrhage
• White cells in casts indicates inflammation of the
kidney pelvis (pyelonephritis)

When WBC>10: we DO Culture of the urine (whether we find

bacteria or not) , When WBC < 10: we DON'T Culture the urine
(But if the sample contains bacteria we culture)
 Procedure for Gram stain:
➢ Bacterial culture section
1- Add Crystal violet for (2 min) and
Processing area then wash off with water.
2- Add Iodine for (1 min) then wash off
Microbiology relies on two important steps for identifying organisms
with water
1) Gram stain 3- Add (Alcohol) for (few second) then
2) Culture wash off with water.
4- Add Safranin for (1-2 min) then wash
For example, Urine culture off with water.
Urine specimens need to be examined within 2 hours. Urine that is left to
Result of Gram stain
stand too long becomes alkaline because bacteria begins to split the urea
contained in urine into ammonia Red color Gram negative Because
cell wall has only a small
We use two types of loops: amount of peptidoglycan.
▪ Larger loop (0.01 mL) It is used for sterile urine because it gives a greater Gram positive Because
Violet
chance of detecting microorganisms (suprapubic aspirate, cytoscopy) cell wall has a large
color amount of peptidoglycan.
▪ Small loop (0.001) is recommended in such cases in order to increase the
resolution and allow quantification of lower concentrations of bacteria.

1- Culture:
• 5% sheep blood agar
• MacConky agar
• Cystine-lactose electrolyte deficient (CLED)
 2- Incubate plates for at least 48 hours at 350C to 370C in air.
3- Interpretations culture
• If the small loop(0.001) is used (one colony equals 1000 CFU\mL)
Count the approximate number of colonies
>10,000 bacteria/mL (insignificant)
10,000-100,000 bacteria/mL (doubtful significance)
<100,000 bacteria/mL (significant)
• If the larger loop(0.01) is used (one colony equals 100 CFU\mL)
any quantity is significant regardless of the number.
Rapid test in this lab

Pregnancy tests H. pylori Antigen Rapid

Test
Sample : urine
Sample : stool
Principle
work by detecting the Principle
presence of the hormone, Release of antibodies to
search for H. pylori Antigen
human chorionic
gonadotropin (hCG). Time : 15 min
Time : 2 min
Cases in K.F.H
1- sample reception 2- Gram stain and
culture
CSF is commonly
obtained aseptically by Gram stain must be done on
lumbar puncture in a CSF sediment, using one
screw-capped container. drop on sterile slide.
Put a drop of the CSF
sample on:
Blood agar(BAP)
Chocolate agar(Choc)
MacConkey ( MAC)

3- Sample preparation 4- Agglutination test

Boil the sample as should be performed to detects

required in attached kit the following pathogen:
If a white color appears • Streptococcus agalactiae
and the SCF becomes egg- • Neisseria meninigidits
like , it indicates the • Streptococcus pneumonia
presence of protein • Hemophilus influanzae type b
• Escherichia coli k1
• Listeria monocytogenes
Agglutination means a positive
 Interpretations
Three tubes send to
1. Biochemistry lab for protein and glucose evaluation
2. Microbiology for sediment
3. Hematology for cell and differential counts

• If the polymorphonuclear (PMN) neutrophils , protein , glucose = bacterial

• If lymphocytes , Negative bacterial and fungal cultures , Normal glucose , protein = viral infections
Thank you