Activity 13 Urine Culture Inoculating Urine with a calibrated loop The number of microorganisms per milliliter recovered on urine culture can aid in the differential diagnosis of urinary tract infection (UTI). Plastic or wire inoculating loop available commercially, have been calibrated to deliver a known volume of liquid when handled correctly, thus enabling the microbiologist to estimate numbers of organism in the original specimen based on colony forming unit (CFU) of growth on cultures. PROCEDURE: 1st day 1. Gently swirl the specimen bottle to mix the urine specimen. 2. Label all plated media with name of patients, clinical specimen used and date. Label at the bottom of plates and not on the cover. 3. Obtain a disposable calibrated loop. 4. Dip the loop straight into the urine specimen so that the loop part is completely covered. Withdraw straight out. See fig. 16-18 5. Inoculate blood agar plate (BAP) as shown in fig.16-19 6. Incubate at 35 – 37˚C for 18 – 24 hours. 7. Dispose of urine sample: a. Pour disinfectant into the specimen and let it stand for about 2 hours before pouring it into a toilet bowl in the rest room. b. Disinfect the empty container with diluted lysol and wrap in a plastic bag. c. Throw it into the yellow garbage can for infectious specimen. 2nd day Colony count: 1. Count the colonies on BAP after incubation. The number of CFU (colony forming unit) is multiplied by its dilution factor. Such as 1000 (if a 0.001 ml loop was used) or by 100 (if a 0.01 ml loop was used) to determine the number of microorganisms per milliliter in the original specimen. 2. Record result. 3. Antimicrobial treatment or other factors may inhibit initial growth, re-incubate plate with no growth or tiny colonies for an additional of 24 hours before discarding plates. 4. Discard the agar in a plastic bag with disinfectant and throw in the garbage for infectious materials. Disinfect the plates before washing.
Activity 13
Urine Culture Inoculating Urine with a calibrated loop
The number of microorganisms per milliliter recovered on urine culture can aid in the differential diagnosis of urinary tract infection (UTI). Plastic or wire inoculating loop available commercially, have been calibrated to deliver a known volume of liquid when handled correctly, thus enabling the microbiologist to estimate numbers of organism in the original specimen based on colony forming unit (CFU) of growth on cultures.
PROCEDURE: 1st day
1. Gently swirl the specimen bottle to mix the urine specimen.
2. Label all plated media with name of patients, clinical specimen used and date. Label at the bottom of plates and not on the cover.
3. Obtain a disposable calibrated loop.
4. Dip the loop straight into the urine specimen so that the loop part is completely covered. Withdraw straight out. See fig. 16-18
5. Inoculate blood agar plate (BAP) as shown in fig.16-19
6. Incubate at 35 – 37˚C for 18 – 24 hours.
7. Dispose of urine sample:
a. Pour disinfectant into the specimen and let it stand for about 2 hours before pouring it into a toilet bowl in the rest room.
b. Disinfect the empty container with diluted lysol and wrap in a plastic bag.
c. Throw it into the yellow garbage can for infectious specimen.
2nd day Colony count:
1. Count the colonies on BAP after incubation. The number of CFU (colony forming unit) is multiplied by its dilution factor. Such as 1000 (if a 0.001 ml loop was used) or by 100 (if a 0.01 ml loop was used) to determine the number of microorganisms per milliliter in the original specimen.
2. Record result.
3. Antimicrobial treatment or other factors may inhibit initial growth, re-incubate plate with no growth or tiny colonies for an additional of 24 hours before discarding plates.
4. Discard the agar in a plastic bag with disinfectant and throw in the garbage for infectious materials. Disinfect the plates before washing.
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