Gram Negative Bacilli Method

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Gram Negative Bacilli Method

A loopful of stool should be inoculated into a plate of deoxycholate citrate agar and tube of selenite and incubate at 35ºC of temperature.

Campylobacters palte should be also inoculated and incubate in 5%CO2 /N2  atmosphere at 43ºC.

Take somw quantity of the stool emulsify a further portion of it with a peptone water, and used Wilson and Blair plate and MacConkey plate inoculate foe enteric fever if suspected and incubate at 35ºC.

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Next examine the plate for any growth of colonies.

Pick a suspected colony and identify through a biochemical reactions, by agglutination test.

Now bring out the selenite  F medium onto a MacConkey agar plate and incubate at 35ºC overnight and examine the colony growth either Non-lactose-fermenting colonies present in the medium.

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When vibrio cholerae is suspected inoculate the specimen into alkaline peptone water and thiosulphate citrate bile salt medium. But if staphylococcus araeus is suspected then inoculate the specimen in a salt cook meath broth or Baird Parker agar and make a Gram film form the inoculation.

Ether Concentration Test:  these method ii used for examination and isolation of mycobacterium tuberculosis prepared a thick saline suspension of stool in a screw-capped bottle.

The same volume of ether should be added and shake very well.

Then centrifuge at 3000rpm for 5 minute the supernatant should be pipette into a clean container, then make a smear the supernatant.

The smear should be stain by Ziehl-Neelsen stain, after that examine with microscope in case, culture is needed take the layer and inoculate into a low-enstain-Jensen  slopes.

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Incubate at 35ºC for a weeks the examination of the culture weekly is guaranteed.

Finally the stool should be treated as if it is a sputum.

How to send fluid to Laboratory

Fluid are message to laboratory in a sterile container, the container must have narrow mouth.

Universal bottle container with 3ml of 3.5% sodium citrate as an anticoagulant is incorporated in other to prevent any clothing of the fluid in case of sample that will cloth when expose to air before it get the laboratory. The same techniques is applicable to CSF. In case of CSF, the fluid should is usually sterile, but sometimes being contaminated either from the point of collection or the container used to collect the blood is not sterile as a result of that introduce contaminant to sample, also contaminant can be introduce during culturing procedure.

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