title: Comparison of three nonradioisotopic polymerase chain reaction-based methods for detection of human immunodeficiency virus type 1.
abstract: Three nonradioisotopic polymerase chain reaction (PCR)-based detection techniques were evaluated for sensitivity and specificity in detecting human immunodeficiency virus type 1 (HIV-1) proviral DNA in peripheral blood mononuclear cells. The Roche prototype HIV-1 PCR assay, the Du Pont enzyme-linked oligonucleotide sandwich assay (ELOSA), and the Gen-Probe hybridization protection assay (HPA) were compared with a standard radioisotopic oligonucleotide solution hybridization (OSH) technique. A panel of 111 well-characterized clinical samples that included peripheral blood mononuclear cells from 48 healthy, low-risk, HIV-1 antibody-negative subjects, 24 antibody-positive subjects with stable CD4 counts of less than 200/mm3, and 39 antibody-positive subjects with stable CD4 counts of greater than 800/mm3 were studied. Each method demonstrated good specificity, ranging between 96 and 100%; those of the OSH and ELOSA (Du Pont) were 100%, those of the HPA (Gen-Probe) were 100% with one probe and 96% with the other probe, and that of the HIV-1 PCR assay (Roche) was 96%. Sensitivities ranged from 96 to 100% for the low-CD4-count group, with the OSH, the HIV-1 PCR assay (Roche), and the HPA (Gen-Probe) all attaining a sensitivity of 100%. For the high-CD4-count group, sensitivities ranged from 69 to 97%, with the OSH attaining a sensitivity of 97% and the HPA attaining sensitivities of 97% with one probe and 95% with the other probe. These data indicate that the nonradioisotopic techniques are sensitive and specific for the detection of HIV-1 proviral DNA in clinical samples.
authors.affiliation: Northwestern University, Chicago, Illinois 60611.
authors.forename: A J
authors.initials: AJ
authors.lastname: Whetsell
authors.forename: J B
authors.initials: JB
authors.lastname: Drew
authors.forename: G
authors.initials: G
authors.lastname: Milman
authors.forename: R
authors.initials: R
authors.lastname: Hoff
authors.forename: E A
authors.initials: EA
authors.lastname: Dragon
authors.forename: K
authors.initials: K
authors.lastname: Adler
authors.forename: J
authors.initials: J
authors.lastname: Hui
authors.forename: P
authors.initials: P
authors.lastname: Otto
authors.forename: P
authors.initials: P
authors.lastname: Gupta
authors.forename: H
authors.initials: H
authors.lastname: Farzadegan
mesh_terms: D001483:Base Sequence; D015342:DNA Probes; D004279:DNA, Viral / Q000097:blood / Q000235:genetics*; D005069:Evaluation Studies as Topic; D015658:HIV Infections / Q000175:diagnosis / Q000382:microbiology; D015497:HIV-1 / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000382:microbiology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D011533:Proviruses / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity
chemical_list: D015342:DNA Probes; D004279:DNA, Viral