Document 2752
 DOCN  M94A2752
 TI    Targetting of HIV-DNA by probe DNA beads to improve PCR sensitivity.
 DT    9412
 AU    Tano H; Fan K; Kitajima M; Kasai K; Hayashi T; Kondo M; Imai M; Japan
       Synthetic Rubber Co., Ltd. Tsukuba Research Laboratory,; Tsukuba.
 SO    Int Conf AIDS. 1994 Aug 7-12;10(1):232 (abstract no. PB0358). Unique
       Identifier : AIDSLINE ICA10/94369823
 AB    OBJECTIVE: The HIV-DNA assay by PCR has been widely used. To increase
       the sensitivity and specificity of conventional PCR method for detection
       of HIV-DNA, latex beads bound with probe-DNA were applied to concentrate
       and isolate the target DNA. METHODS: Probe-DNA against HIV-DNA was
       covalently bound to latex bead with carboxylate groups on its surface.
       This latex suspension was hybridized with a model specimen containing
       several copies of HIV DNA in the volume of 30ml, and thereafter
       concentrated to 30 microliters (1000 fold concentrated), by
       centrifugation. Concentrated HIV DNA was used as the template for nested
       PCR detection. RESULTS: Concentrated target DNA was successfully
       detected by PCR, whereas the same specimen without concentration is
       undetectable by PCR. The assay sensitivity was improved in the order of
       100 to 1000. DISCUSSION AND CONCLUSIONS: In this study, it is
       experimentally confirmed that probe-DNA bound latex bead could target
       HIV-DNA to improve PCR assay sensitivity and reduce the nonspecific
       bands. It can be applied for detection of HIV-DNA by PCR after pooling a
       number of individual specimen into one with a larger scale, followed by
       the concentration of the target DNA. In this way, a more efficient and
       lower cost PCR diagnotics can be expected.
 DE    *DNA Probes  DNA, Viral/*ANALYSIS  HIV/GENETICS/*ISOLATION & PURIF
       Latex  Microspheres  Polymerase Chain Reaction/INSTRUMENTATION/*METHODS
       Sensitivity and Specificity  MEETING ABSTRACT

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       protected by Copyright Law (Title 17, U.S.Code).