identification and evaluation of new primer sets for the detection of lentivirus proviral dna.

NLM AIDSLINE Identification and evaluation of new primer sets for the detection of lentivirus proviral DNA. Gelman IH; Zhang J; Hailman E; Hanafusa H; Morse SS; Department of
May 30, 1993

AIDS Res Hum Retroviruses. 1992 Dec;8(12):1981-9. Unique Identifier : We have developed sets of degenerate oligonucleotides designed to detect pol gene sequences from any member of the lentivirus subfamily when used as primers in amplification techniques such as the polymerase chain reaction (PCR). This pan-lentivirus-specific primer set (PLSPS) consists of primers, LV1, LV2, and LV3, based on conserved regions common to lentiviruses only. Our protocol is based on primary amplification with LV1 and LV2 followed by secondary amplification with a nested primer set based on the YM/VDD motif found in all reverse transcriptases (or DDMY, in the opposite direction), and LV3, a block of lentivirus homology nested just downstream of LV1. PLSPS-PCR analysis of DNA from cells infected with HIV-1, HIV-2, SIVmac239, BIV, visna, EIAV, CAEV, OPPV, or FIV resulted in the amplification of appropriately sized products. Sequence analysis of the LV1/2 products, cloned into pBluescript (pBS), indicated that at least 20% (most often, > 80%) contained the predicted lentivirus pol sequence. Greater than 95% of the LV3/DDMY products contained the expected lentiviral sequences. Using the PLSPS, lentivirus pol sequences could typically be detected at levels of one copy in 2 x 10(6) cells after secondary amplification. No specific lentiviral PCR products were detected in DNA from uninfected human or mouse monocytes, feline or bovine leukocytes, mouse, rat or human fibroblast cell lines, chicken embryo fibroblasts, Tahr lung cells, or cell lines infected with the following retroviruses which are not lentiviruses: Rous sarcoma virus, Moloney leukemia virus or Kirsten sarcoma virus, mouse mammary tumor virus, human T-cell lymphotropic virus I, and feline leukemia virus.(ABSTRACT TRUNCATED AT 250 WORDS) Amino Acid Sequence Animal Base Sequence Cell Line Comparative Study DNA, Viral/GENETICS Evaluation Studies Genes, pol Human Lentivirus/GENETICS Molecular Probe Techniques Molecular Sequence Data Oligonucleotide Probes Polymerase Chain Reaction Proviruses/GENETICS Retroviridae/GENETICS Sequence Homology, Amino Acid Species Specificity Support, Non-U.S. Gov't Support, U.S. Gov't, P.H.S. JOURNAL ARTICLE

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Information in this article was accurate in May 30, 1993. The state of the art may have changed since the publication date. This material is designed to support, not replace, the relationship that exists between you and your doctor. Always discuss treatment options with a doctor who specializes in treating HIV.