Detection of Human Immunodeficiency Virus-1 Nucleic Acid on Inactivated Filter Paper Disks by Polymerase Chain Reaction and Microtiter Plate Assay

J-STAGE
  • Kunisada Takao
    Division of AIDS Virus, AIDS Research Center, National Institute of Health
  • Ando Shuji
    Division of AIDS Virus, AIDS Research Center, National Institute of Health
  • Saito Kunihiro
    Department of Clinical Nutrition, Tokyo University of Agriculture
  • Eshita Yuki
    Division of AIDS Virus, AIDS Research Center, National Institute of Health
  • Röder Wolfgang
    Abteilung für Allgemein- und Unfallchirurgie
  • Kruse Michael
    Institut für Physiologische Chemie, Abteilung "Angewandte Molekularbiologie," Universität
  • Müller Werner E. G.
    Institut für Physiologische Chemie, Abteilung "Angewandte Molekularbiologie," Universität
  • Ushijima Hiroshi
    Division of AIDS Virus, AIDS Research Center, National Institute of Health

Abstract

Human immunodeficiency virus type 1 (HIV-1) in cultured cells, peripheral blood samples and sera were adsorbed on filter paper disks and inactivated by heat or ethanol. Two procedures, the polymerase chain reaction (PCR) and microtiter plate assay (HMPA) were used to detect the nucleic acid. The sensitivity after different heat treatments with nested PCR for HIV-1 DNA (or nested reverse transcription-PCR for HIV-1 RNA) was identical regardless of whether the samples were examined immediately or one month later. Inactivation by ethanol treatment resulted in a slight loss of sensitivity. The HMPA proved to be as reliable and specific as the conventional PCR technique. We conclude that the heat-treated filter paper disk assay is suitable for identifying HIV nucleic acid in clinical samples sent to the laboratory from a distance, e.g. in an envelope.

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