Article Abstract:
Sufficient concentration of Triton X-100 was observed to improve the immunomagnetic separation of Coxiella burnetii from milk. PCR-enzyme-linked immunosorbent assay (PCR-ELISA) can detect coxiellas more effectively than the standard PCR. PCR-ELISA is therefore believed to be applicable for the simultaneous assay of huge number of samples. However, the number of cows from which raw milk tested positive for coxiellas by PCR-ELISA failed to agree with that found with the antibody to coxiella by indirect immunofluorescence assay. This is due to the differences in the infectious route, infectious dose, or the timing of yielding the antibody and the period of duration of the antibody.
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Article Abstract:
A sample preparation method that uses silica binding of DNA for detecting polymerase reaction chain (PCR) of Coxiella burnetii from different clinical specimens is presented. The method was tested on more that 600 milk samples from 21 cows that carry the C. burnetii agent and have fertility problems. Results show that the method can detect C. burnetii in naturally infected specimens. However, the sensitivity of the method is inferior to other PCR-based detection methods.
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Article Abstract:
An alternative cloning method is developed, which involves excision of individual Coxiella burnetii-laden parasitophorous vacuole (PV) from infected cell monolayers by micromanipulation. It is found that micromanipulation is an efficient and reproducible procedure for obtaining Coxiella burnetii clones.
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