HIV病毒库分子检测方法和临床研究进展

Molecular detection methods and clinical research progress of HIV reservoir

抗逆转录病毒治疗(ART)可有效降低病毒复制速度,控制病毒数量,但是不能彻底清除人类免疫缺陷病毒,主要原因是病毒库的存在。病毒库的存在及其大小与艾滋病疾病进展、抗病毒治疗效果、病毒反弹等密切相关,但是病毒库的检测一直是个难点。Siliciano等于1995年报道了定量病毒生长测定法,并成为公认的经典方法。但是由于该法需要大量血浆和体外培养的P3实验室条件而无法在多数实验室中进行。各个实验室建立了自己的方法,导致各实验室数据没有可比性。最近报道的实时荧光定量聚合链酶式反应可以同时检测总DNA、整合DNA和环DNA,与经典方法的一致性高,有望成为一种通用的敏感快速的病毒库检测方法。本综述将重点介绍经典的定量病毒生长测定法、实时荧光定量聚合链酶式反应和滴液数字聚合链酶式反应分析方法在检测HIV病毒库方面的应用以及每种方法的优点和局限性。

Antiretroviral therapy(ART)can effectively reduce the rate of viral replication and control the amount of the human immunodeficiency virus(HIV),but it cannot completely eliminate the virus due to the presence of viral reservoirs.Siliciano et al.reported a quantitative virus growth assay in 1995,which has become a recognized classic method.However,due to the need for large amounts of plasma and P3 labora-tory conditions for in vitro cultivation,it cannot be conducted in most laboratories.As a result,each laboratory has developed its own methods,leading to a lack of comparability between data from different laboratories.Re-cently,a real-time fluorescence quantitative polymerase chain reaction has been reported,which can simultane-ously detect total DNA,integrated DNA,and circular DNA with high consistency compared to classic meth-ods.This new method is expected to become a universal sensitive and rapid virus library detection method.This review will focus on the application of classical quantitative virus growth determination,real-time-quantitative polymerase chain reaction,and droplet digital polymerase chain reaction analysis methods in the detection of HIV virus libraries,as well as the advantages and limitations of each method.