Introduction

1　Introduction

Human immunodeficiency virus type 1（HIV-1）is the etiological agent of the acquired immunodeficiency syndrome（AIDS）.The accurate and quantitative determination of HIV-1 in the human host is essential for the diagnosis of HIV-1 infection and for the assessment of the efficacy of both antiviral therapy and therapeutic vaccine intervention.In addition，the evaluation of the copy number of HIV-1 RNA sequences in specimens collected for clinical trials requires extensive handling，shipping and storage.An early study showed that cell-free infectious virus in plasma is an indicator of HIV-1 replication（Zagury et al.，1985）.Previous studies with quantitative plasma culture indicated that plasma viremia is a useful marker of disease progression and is a potential marker for evaluating the response to antiviral chemotherapy（Ho et al.，1989；Coombset al.，1989）.Viral culture，although sensitive，is cumbersome，timeconsuming and subject to selection of viral strains.The polymerase chain reaction（PCR），which specifically amplifies HIV nucleic acids from a clinical specimen，provides a useful tool for the direct detection of HIV-1 sequences（Ou et al.，1988，1990；Hewlett et al.，1988；Rogers et al.，1989；Schnittman et al.，1989；Genesca et al.，1990；Jackson et al.，1990；Bagnarelli et al.，1991；Daar et al.，1991；Holodniy et al.，1991；Ottmann et al.，1991；Michael et al.，1992；Saksela et al.，1994）.A sensitive assay was developed to measure quantitatively small amounts of HIV-1 RNA directly from plasma.Ten copies of HIV-1 RNA can be detected by RT-PCR amplification and solution oligonucleotide hybridization detection.However，for quantitative results，the most satisfactory range is 50—1，000 copies.With this assay，HIV RNA is extracted with guanidinium thiocyanate and quantitated relative to a full length synthetic HIV-1 cRNA standard.The inter-assay variation among identical replicates is within a two-fold difference.Using this assay，we have evaluated the stability of HIV-1 in plasma，the effect of plasma constituents from different individuals on the assay，and the distribution of HIV RNA sequences in whole blood of seropositive patients.

HRI，Research Inc.，2341 Stanwell Dr.，Concord，CA 94520，USA

Accepted 5 October 1994(https://www.daowen.com)

此文发表于1995年“J.Virol.Methods”52：287-299.