Tuesday, July 17, 2007

urinalysis/routineµbiology

Hi~liuqian here~
First of all ,I am sorry abt the late posting!in my attachment place , we have chance to go every lab and rotate per 2 weeks. This is the 3rd week already ,so I ve already attached to 2 labs, urinalysis/ non-routine and micro lab.
First, I say something about the urinalysis/ non-routine. The tests running there are urine FEME, CSF analysis, urine osmolality and urine phase contrast.

URINE FEME:
This is the most common test running in this lab. It is ordered by doctor for urinary tract infection.
Procedure:
A: chemical testing of urine analytes
1. Mix the urine to be tested by inverting the sample several times before opening the sample cap
2. Completely immerse al reagent areas of the strip
3. Record the result of PH, Albumin, Sugar, Ketone, and blood.
B: microscopic examination
Microscopic examination of urine sediment is examined unstained.
1. Mix the urine sample and transfer to the KOVA glasstic slide chamber with plastic pipette
2. Quantitate and examine all cells under microscope
3. For low cell count samples count the total cells of 4 complete quadrants of the
counting rids
4. For high cell count samples count 10 small grids within different quadrant

CSF FLUID
This test is ordered by doctor for meningitis.
The things we are looking for in CSF analysis:
1. appearance
2. cell counts with differential
3. glucose
4. total protein
5. smears for Gram’s stain and AFB stain
6. Indian ink preparation for encapsulated crytococcus

URINE PHASE CONTRAST
Urine phase contrast is a method of distinguish glomerular bleeding from causes of haematuria by examining the appearance of RBC in urine.
Procedure:
1. centrifuge 10 or 12 ml of urine for 10 mins at 2000rpm
2. discard 9 or 11 ml of supernatant
3. resuspend 1 ml of sediment and charge into the KOVA chamber
4. examine for cast, epithelial cells, micro-organism, crystals at200X
5. quantitate casts, EC, RBC and WBC at 400 X
6. roughly calculate the ratio of ISO to DIS(ISO RBC is very shine and round under the phase contrast dark background microscope while DIS RBC is dark and variable in shape.

I attached to micro-lab after 2 weeks in urine bench. Basically doctor sent specimen to micro-lab to find out which bacteria causes the infection. So there are different types of specimen, urine, blood, fluid etc.
The specimens are divided into stool, urine, and blood and miscellaneous.
STOOL for food handler------selenite broth------after 16-18h 35 dgree-------salmonella-shigella plate
sceen for fecal pathogen---salmonella-shigella plate+ MacConkey plate+Campylobater plate
Note: if watery stool.must +blood agar+ alkaline peptone+triosulphate citrate bile salt sucrose agar----------------subculture alkaline peptone water to thiosulphate citrate bile sucrose agar

Load the BLOOD vials into BACTEC
Blood specimen normally sent in vials, aerobic or anaerobic. The media contained in the vials consist of soybean casein digest broth with resin and 0.05% w/v sodium polyenrthol sulphate(SPS). Bactec culture vials that are equipped with a sensor for the detection of CO2. Growth of microorganism within the vial results in the production of CO2, which reacts with a dye in the sensor. Fluorescence of the sensor proportional to the CO2 production is measured by a photo-detector. Vial that exeed a pre-determined threshold level will be flagged by the BACTEC instrument as “positive”.
STERILE FLUID:
Blood agar Blood agar
+ Chocolate agar +MacConkey agar +
CDC anaerobic agar + Cooked meat
&
Cooked meat must be
Subcultured after 24h
Incubation
Subculture into chocolate agar
+ CDC anaerobic agar
NON-STERILE FLUID:
Blood agar + Chocolate agar Chocolate agar + MacConkey agar
Finally i finish writing . Enjoy reading:)
Liu Qian
0503935i
TG02

21 comments:

ALsubs said...

Hi Liu Qian! U mentioned that u dip the test strip into urine to test for ketones right? Under what circumstances will ketones be found the urine?

Shu Hui
TG02

Distinction in Disaster! said...
This comment has been removed by the author.
Distinction in Disaster! said...

hello! would like to ask abt the CSF fluid test for meningitis.. based on the parameters u pointed out, wad are the normal ranges and the abnormal ranges?

Joan
TG01

ALsubs said...

hey~Joan
*normal appearance is clear
*the normal white blood cells are between 0 and 5
the normal red blood cell count is 0
*The glucose levels should be between 50 to 80 mg/100 ml. Levels may vary slightly depending on the amount of glucose in the blood, however, the level in the CSF should be about 2/3 the level in the blood.
*The normal protein range varies from lab to lab, but is typically about 15 to 45 mg/dL.
*CSF is sterile fluid in the body ,so there should not have microorganism


LIU QIAN:)

ALsubs said...

Hi!shuhui~
when not enough insulin is present (or, if a person is resistant to insulin as in the case of some with type 2 diabetes and other metabolic disorders) glucose does not move into the cells and can build up in the bloodstream. The body senses cells are not being fed it will turn to fat stores and lean muscle mass as a source of energy.burning of fat wil produce ketone.So a positive test may indicate Metabolic abnormalities, including uncontrolled diabetes or glycogen storage disease.

LIU QIAN

MedBankers said...

Hellooo... wa.. it sounds so tough to work in the microbio bench. By the way, do you do test for Acid Fast Bacilli (MAS is the test of the name but i not sure what the full term is), a smear and sometimes plus a culture using sputum?

-Pei Shan- TG02

ALsubs said...

yo Liu Qian =D

one question here... how does the results of PH, Albumin, Sugar, Ketone, and blood aids in determining urinary tract infection?

Dorene =D
TG02

Distinction in Disaster! said...

Hello! =DD

Since there is a need to do a chemical testing of the urine analytes, how would the results of pH, Albumin, Glucose and Ketone change accordingly when tested on a patient with UTI? Thanks!

Charmaine Tan
TG01

Star team said...

hello. Urine phase contrast is also done in my lab. What is done if you are unable to identify the cells? (e.g too many cells until the whole field is blur)

Also, for FEME tests, is fluid from other parts of the body (e.g pleural or knee aspirate) conducted for tests as well?

Martin
TG02

Star team said...

hello, u mentioned that blood samples are sent in vials that contain media (soybean casein digest broth with resin and 0.05% w/v sodium polyenrthol sulphate(SPS)).

What is the purpose of this media?

Jo-anne Loh
TG02

Distinction in Disaster! said...

Hey, a question here

What is ISO and DIS RBC? and why must their ratio be calculated for Urine phase contrast?

Adrian TG01

ALsubs said...

Hellooo... wa.. it sounds so tough to work in the microbio bench. By the way, do you do test for Acid Fast Bacilli (MAS is the test of the name but i not sure what the full term is), a smear and sometimes plus a culture using sputum?

-Pei Shan- TG02

yes, we do the acid fast test stain for mycobacteria which is TB.
liu qian :)

ALsubs said...
This comment has been removed by the author.
ALsubs said...

hi dorene&Charmaine

the ph, albumun, sugar, krtone, and blood is not for UTI, but help the doctor to determine the function of the kidney~

liu qian:)

ALsubs said...

hello martin~
for the urine ,they just report as >2250
for other fluid ,they only look on the white blood cells, if too many white blood cells to see WBC, they do a 10X dilution of the specimen ,then count the WBC,report as the number times 10

Liu Qian :)

ALsubs said...

hello Jo-anne!
i guess the media is the base for bacteria to grow~

Liu Qian :)

ALsubs said...

hi Adrian
Dysmorphic RBCs have irregular membrane blebs or shapes, often with reduction in hemoglobin. The presence of more than 80% dysmorphic cells in a sample is highly suggestive of a glomerular disease, whereas the presence of less than 20% dysmorphic cells is associated with a urologic source.

Liu Qian:)

ALsubs said...

Hi liu qian!!

when u mentioned abt STOOL for food handler-->selenite broth-->salmonella-shigella plate

why do u use selenite brothsalmonella shigella plate???

Vinodhini
TGO2

VASTYJ said...

hi, sry for late comments.. i juz wanna check.. for total protein in ur CSF.. do u read from the refractometer or do u do it on machines with chemical reactions?

Jia Hao

ALsubs said...

hi vino~
because for the food handler. they forcus on screening of samonella and shigella.


Liu Qian :)

ALsubs said...

hi jia hao~
for the total protein of CSF, we do on the machine in the biochemustry section~

Liu Qian :)