[Prior enrichment of HIV-DNA with probe-DNA particles for an efficient PCR diagnosis].
PCR mediated detection of HIV-DNA has been widely used. However, compared with traditional immunological diagnoses, the extraction of DNA is a laborious and time consuming step. We have developed a procedure for the efficient isolation and concentration of HIV-DNA from cell lysates. We report here a novel method by one can recover HIV-DNA in a small volume (approximately 50 microliters) of solution from a large volume of crude cell lysate which contains as few as several copies. The method uses the specific hybridization of HIV-DNA to HIV probe-DNA particles. This prior enrichment augmented the sensitivity in the detection of HIV-DNA by PCR, and allows us to make a diagnosis even if the specimen contained an extremely low copy number of HIV-DNA molecules in a large volume, which would have otherwise resulted in false-negative data with the conventional extraction method. The method also enables the examination of 100 individual blood specimens in a combined form. Thus, the application of the present enrichment procedure with HIV probe-DNA particles should reduce the labor and cost of HIV diagnosis, since the HIV positive samples represent a very minor group of people among specimens subjected to clinical laboratory tests, and particularly, among blood samples voluntarily donated to be used for transfusions.
Fan
K
K
Japan Synthetic Rubber Co., Ltd. Tsukuba Research Laboratory.
Tano
H
H
Kitajima
M
M
Tamatsukuri
S
S
Furuichi
Y
Y
Hayashi
T
T
Imai
M
M
D001483:Base Sequence; D002498:Centrifugation; D017931:DNA Primers; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D006678:HIV / Q000235:genetics*; D015658:HIV Infections / Q000175:diagnosis; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D015342:DNA Probes; D004279:DNA, Viral