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Virologica Sinica, 16 (2) : 119-123, 2001
Research Article
Establishment of Fluorescent Real Time Quantitative PCR for Detecting HIV-1 and Its Application
Kunming Institute ofZoology,Chinese Academy ofSciences.Kunming 650223.China
Accurate determination of HIV一1 pmviral burden and viralload is very useful in prognosis of
HIV一1 infected patients and in assessment of drug for therapy of AIDS patients.In order to establish a
quantitative method in detecting HIV一1 proviral burden and viralload.8E5 cell line and a recombinant
RNA constructs were used as the HIV一1 proviral DNA and viral RNA external references,respective—
ly The PCR products were Labeled with the fluorescent D A dye SYBR green.The amount of burden
or load was measured by GeneAmp 5700 Sequence Detection System Using this method.the HIV 1
pmvlral burdens in PBMC of patient and in cell suspension treated with the compounds AZT.GL and
W T were measured.HIv_1 viraIloads in supernatant of the cell culture treated with the above com—
pounds were also determined.The therapeutic indices(TIs)of the compounds calculated based on the
inhibition of virus induced syncytlal formation.and inhibitionn of proviral burdens and viralloads were
compared,and their TIs successively increased.The fluorescent real time quantitative PCR poseses
very good specificity.sensitivity and duplication.TI value of a drug based on inhibition of proviral
burden in cell culture,and the proviral burden in PBMC of patient may be useful in evaluating a drug
on eradicating provirus from resting and memory CD4 T cells
  Published online: 5 Jun 2001
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