Detection of @entity5092 within infected XXXX tissues by using a hand-held PCR thermocycler.

@entity1 @entity19 @entity5092 @entity281 @entity1576 @entity6588
The diagnosis of @entity1 cases of @entity6588 often relies upon the demonstration of an antibody response to @entity5092 or the direct culturing of the bacteria from the patient. Antibody response is not detectable until 2 weeks or more after @entity281 , and culturing requires special media and suspicion of @entity6588 . In addition, handling live @entity5092 poses a risk to laboratory personnel due to the highly infectious nature of this pathogen. In an effort to develop a rapid diagnostic assay for @entity6588 , we investigated the use of TaqMan 5' hydrolysis fluorogenic PCR to detect the organism in tissues of infected @entity19 . @entity19 were infected to produce @entity1576 . The fopA and tul4 genes of @entity5092 were amplified from infected spleen, lung, liver, and kidney tissues sampled over a 5-day period. The samples were analyzed using the laboratory-based Applied Biosystems International 7900 and the Smiths Detection-Edgewood BioSeeq, a hand-held portable fluorescence thermocycler designed for use in the field. A comparison of culturing and PCR for detection of bacteria in infected tissues shows that culturing was more sensitive than PCR. However, the results for culture take 72 h, whereas PCR results were available within 4 h. PCR was able to detect @entity281 in all the tissues tested. Lung tissue showed the earliest response at 2 days when tested with the ABI 7900 and in 3 days when tested with the BioSeeq. The results were in agreement between the ABI 7900 and the BioSeeq when presented with the same sample. Template preparation may account for the loss of sensitivity compared to culturing techniques. The hand-held BioSeeq thermocycler shows promise as an expedient means of forward diagnosis of @entity281 in the field.
