Evaluation of a new non-isotopic sandwich hybridization for the detection of HIV1-specific PCR products.
A new non-isotopic sandwich hybridization assay was developed to detect human immunodeficiency virus type 1 (HIV1) provirus amplified by the polymerase chain reaction. The sensitivity and specificity of this new technique using 96-well microplates as the support for the sandwich hybridization procedure and stable enzyme-linked oligomer as the detection probe were compared with those of Southern hybridization using a 32P-labelled oligomer probe. Three laboratories studied 437 peripheral blood mononuclear cell samples from 294 different subjects including both HIV1-seropositive and -seronegative individuals. The non-isotopic assay exhibited a sensitivity of 99.5% and a specificity of 99.1% when compared with the Southern procedure. In addition, the non-isotopic assay gives clear numeric data, is safe when handling, and is especially adapted to large-scale analyses.
Aubin
J T
JT
Laboratoire de Bactériologie-Virologie, CNRS EP 57, C.E.R.V.I., Hôpital Pitié-Salpêtrière, Paris.
Boulay
D
D
Chapus
A
A
Bréchot
C
C
Laure
F
F
Agut
H
H
D001483:Base Sequence; D015139:Blotting, Southern; D004279:DNA, Viral / Q000032:analysis*; D005069:Evaluation Studies as Topic; D015751:Genes, pol; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007554:Isotopes; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D011533:Proviruses / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity
D004279:DNA, Viral; D007554:Isotopes