| 摘要 |
<p>The present invention provides a method for detecting a target nucleic acid fragment in a clinical specimen. A sample of the specimen is solubilized and treated to denature the nucleic acids therein. The sample is contacted with at least one probe complex comprising a sequence complementary to a portion of the target fragment, as well as a first member of a specific binding pair. Following hybridization of probe and target, the probe complex is contacted with a solid substrate linked to the second member of the specific binding pair to isolate hybridized target fragment in a probe-target-solid substrate ternary complex. The isolated ternary complex is separated from the sample, and the target fragment and probe complex are released into solution. The released target fragment is amplified by PCR or RT-PCR, and the presence of the target fragment in the clinical specimen is detected. The method may be used on a variety of specimen types, and is useful for detecting pathogens such as Borrelia burgdorferi and species of Babesia. Specific chaeotropic salt solutions, wash buffers, and conditions for amplification are disclosed. Preferred probe complexes and primers for the detection of Borrelia and Babesia pathogens are disclosed.</p> |
