Protocols

Fertilizer reaction experiments

Summary

Learn how to operate the Fertilizer Reaction and acquire a preliminary knowledge of its result analysis.

Operation method

quantitative agglutination test

Principle

With known typhoid H, O antigen and A, B paratyphoid H antigen, and the patient's serum for quantitative agglutination test, in order to determine the presence or absence of the corresponding antibody in the patient's serum, according to the amount of antibody content and the growth of the situation, can help to diagnose typhoid fever and paratyphoid fever.

Move

1. Take 28 small test tubes and place them in four rows of 7 tubes each on a test tube rack, labeling the first tube in each row with the symbols H, O, PA, and PB.

2. Add 0.5 mL of saline into each tube.

3. Add 0.5 ml of 1:10 patient serum to the first tube of each row and mix well. Aspirate 0.5 ml from the first tube of the first row.

Inject 0.5 ml into tube 2 of the first row for doubling dilution, dilute sequentially to tube 6, and discard 0.5 ml from tube 6, with tube 7 as the control.

4. The other three rows are diluted in the same way.

5. Starting from tube 7 in each row, add bacterial solution to each tube from back to front. Add 0.5 ml of H bacterial solution to each tube in the first row, 0.5 ml of bacterial solution to each tube in the second row, 0.5 ml of PA bacterial solution to each tube in the third row, and 0.5 ml of PB bacterial solution to each tube and in the fourth row.

6. Shake the test tube rack, mix the above liquids, placed in 37 ℃ 18 ~ 24 h after taking out to observe the results.

7. Result observation

Do not shake the test tube to avoid dispersion of clots. Look at the control tube first, then the test tube.

The degree of agglutination is indicated by "+", "++", "+++", "++++", "++++", and the degree of non-agglutination is indicated by "1". ++++": complete agglutination, the bottom of the tube has a membrane-like agglutination block with irregular edges, and the upper layer of liquid is clear and transparent.

"++++": most of the agglutination, the clots are not all sunk in the bottom of the tube but slightly spread, the upper layer of liquid is clear.

"++": partial agglutination, some agglutinated clots at the bottom of the tube, the upper liquid layer is slightly turbid.

"+": a small part of the agglutination, the bottom of the tube has a small amount of agglutinated pieces, the upper layer of liquid is more turbid.

"I": no agglutination, the liquid is uniformly turbid.

Record the results of the experiment and the agglutination efficiency.



Common Problems

1. How to isolate and identify dysentery bacilli from the feces of patients with acute bacillary dysentery?

2. Why are the four antigens "H", "O", "PA" and "PB" used in the Fidelity reaction?

3. how do you analyze the results of fiducial reactions in your laboratory?


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Categories: Protocols
Explore topics: Microbiology experiment

Da — when not otherwise indicated, molecular weight units are daltons.   Mw — weight-average molecular weight.   Mn — number-average molecular weight.

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Cite this article

Aladdin Scientific. "Fertilizer reaction experiments" Aladdin Knowledge Base, updated Dec 24, 2024. https://www.aladdinsci.com/us_en/faqs/fertilizer-reaction-experiments-en.html
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