Document 0718
 DOCN  M95A0718
 TI    Detection of reverse transcriptase activity by enzyme-linked
       immunosorbent assay in human immunodeficiency virus type 1.
 DT    9510
 AU    Tomita N; Miyahara M; Satoh H; Suzuki K; Kitajima K; Miyamoto K; Okayama
       Red Cross Blood Center, Japan.
 SO    Acta Med Okayama. 1995 Apr;49(2):69-73. Unique Identifier : AIDSLINE
       MED/95343780
 AB    An enzyme-linked immunosorbent assay (ELISA) using biotin-labelled
       oligo-dT primer and digoxigenin (Dig)-dUTP was designed to measure the
       reverse transcriptase (RT) activity of human immunodeficiency virus type
       1 (HIV-1). The ELISA system involves the selective detection step of a
       newly synthesized cDNA by two specific bindings, biotin-streptavidin
       binding and alkaline phosphatase (AP)-conjugated anti-Dig-Dig binding,
       and the enzymatic amplification step to increase coloring generated by
       AP. This method was used to measure the activity of RT in the culture
       supernatants of peripheral leukocytes obtained from four
       anti-HIV-1-positive persons cocultivated with those from four
       anti-HIV-1-negative persons. RT activity was detected in all of four
       anti-HIV-1-positive culture supernatants but not in those cultivated
       with anti-HIV-1-negative supernatants alone. Thus, our improved ELISA
       for detection of HIV-1 appears to be sensitive enough and useful for
       routine laboratory work. This non-radioactive method will also be useful
       for detecting other retroviruses and for screening of RT inhibitors.
 DE    Cells, Cultured  DNA, Complementary/ANALYSIS  DNA, Viral/ANALYSIS
       Enzyme-Linked Immunosorbent Assay/*METHODS  Human  HIV Core Protein
       p24/ANALYSIS  HIV-1/*ENZYMOLOGY/ISOLATION & PURIF  Leukocytes/VIROLOGY
       Reverse Transcriptase/*ANALYSIS  Sensitivity and Specificity  JOURNAL
       ARTICLE

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