摘要:

AbstractThe clinical performance of a modified polymerase chain reaction (PCR) testing algorithm was evaluated for confirming the presence of HIV‐1 proviral DNA in peripheral blood mononuclear cells. A whole cell lysate, rather than phenol‐purified DNA, was used for PCR amplification, under systematically optimized conditions designed and verified within each PCR run to detect as few as 10 copies of proviral DNA. A sequential testing algorithm was designed requiring reactivity in duplicate (with corresponding non‐reactivity in negative controls) with at least two sets of primers, before reporting a specimen as HIV‐l‐positive. In 196 specimens from patients staged according to the Walter Reed staging system, the PCR test sensitivity and the coculture isolation rate (in parentheses) were found to be: 97% (71%), 100% (85%), and 100% (76%) in stage 1, stage 2, and stage 3 specimens, respectively; and 100% (100%) in stage 4, 5, and 6 specimens. Results were uniformly negative for PCR and coculture isolation from 21 blind negative specimens and 105 (negative) donor leukopacks. These data indicate that this PCR testing algorithm is more accurate than tissue culture isolation methods, especially with early stage patients, and results in detection of HIV‐1 in virtually 100% of seropositive individuals, with no fals

ISSN:0146-6615    

DOI:10.1002/jmv.1890330112

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1991

数据来源: WILEY

摘要:

AbstractThis is the first report of the direct detection of HTLV‐I RNA in uncultured peripheral blood mononuclear cells (PBMNC's) of patients with tropical spastic paraparesis and HTLV‐l‐associated myelopathy (TSP/HAM) and their spouses, using the technique of in situ hybridization. Twenty‐one Colombian patients were tested, all of whom had antibodies to HTLV‐I; the presence of HTLV‐I proviral DNA in their PBMNC's was confirmed by the polymerase chain reaction technique. Of the 21 patients 15 had a clinical diagnosis of tropical spastic paraparesis (TSP/HAM), 5 were asymptomatic relatives, and 1 patient had leukemia. In situ hybridization was positive in samples from 5 patients; 2 of these were TSP/HAM patients and the other 3 were healthy wives of TSP/HAM patients. This study demonstrates for the first time that viral RNA is expressed in uncultured PBMNC's of some patients with TSP/HAM in whom proviral DNA is also present; furthermore, the detection of HTLV‐I RNA in the blood of female partners of TSP/HAM patients clearly illustrates the high likelihood of HTLV‐I transmission through

ISSN:0146-6615    

DOI:10.1002/jmv.1890330113

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1991

数据来源: WILEY

ISSN:0146-6615    

DOI:10.1002/jmv.1890330101

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1991

数据来源: WILEY