一种简易的HIV-1病毒载量定量筛查方法的建立

摘要

Objective Based on real-time PCR,to develop a simple and feasible method for quantitative HIV-1 viral load screening.Methods A total of 36 plasma samples (18 HIV positive and 18 HIV negative) were collected in Nanning city,Guangxi.RNA was extracted from these samples and then used to PCR amplification with primer LTR-1/ LTR-2.HIV-1 viral loads of these samples were measured by Nuclisens EasyQ HIV-1 v1.2.Correlation between Ct values of RT-PCR and viral loads was analyzed by regression analysis,and then a linear regression equation was obtained.Results The Ct values varied between 29.103-31.610 cycles among HIV-1-infected cases,while negative controls failed to be amplified by RT-PCR.The results show that there is a good linear relationship between Ct values and the logarithm of viral loads (r =-0.51,P =0.03).The regression equation was logY (viral road) =57.55-1.56 × (Ct value).Conclusion Based on real-time PCR,a simple and low cost method for quantitative HIV-1 viral load screening was established.The method has a good consistency with current commercial products and can be used for quantitative screening of HIV-1 infected patients.%目的 建立一种基于RT-PCR法的简易的HIV-1感染者或艾滋病患者血浆病毒载量筛查方法.方法 在广西南宁市采集18例HIV-1感染者和18例阴性对照者血液标本,提取RNA,以引物LTR-1/LTR-2对待测样本进行PCR扩增,获得Ct值.同时,应用Nuclisens EasyQ HIV-1 v1.2法作为对照对这些样本进行病毒载量检测.对Ct值与病毒载量之间进行相关性分析,获得相应的回归方程.结果 待测样本中HIV-1感染者Ct值在29.103～31.610个循环之间,阴性对照者未能扩增出产物.RT-PCR法Ct值与Nuclisens EasyQ HIV-1 v1.2法检测出的病毒载量对数值之间具有良好的线性关系(r=-0.51,P=0.03),回归方程为logY(viral road)=57.55-1.56×(Ct value).结论 所建立的HIV-1病毒载量RT-PCR检测方法操作简单,价格便宜,与商业化试剂盒具有较好的一致性,可用于HIV-1感染定量筛查.