[一种用于快速分型广西流行的HIV-1毒株的巢式多重PCR检测方法]
[A nested multiplex PCR assay for rapid subtyping the prevailing HIV-1 strains in Guangxi].
作者信息
Su Qi-jian, Zhou Ping, Bi Zhi-you, Xiao Xin, Wu Shu-zhi, Cen Ping, Deng Wei, Shao Yi-ming, Liang Hao
机构信息
Center for AIDS Research & School of Public Health, Guangxi Medical University, Nanning, China.
出版信息
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2009 Dec;23(6):482-4.
OBJECTIVE
To establish a rapid nested multiplex PCR assay for subtyping HIV-1 CRF01_AE, CRF07_BC, CRF08_BC, B, and C strains prevailing in Guangxi.
METHOD
Subtype-specific primers were designed for these subtypes based on their gag sequences. The subtypes of HIV-1 samples from Guangxi were determined by nested multiplex PCR and DNA sequencing and phylogenetic analysis, respectively, and then the sensitivity and the specificity of nested multiplex PCR were calculated.
RESULTS
Nested multiplex PCR could correctly classify the 5 known-subtype samples, and were not reactive to all HIV-negative samples. Of the 72 HIV-positive samples, 66 were correctly identified as CRF01_AE, CRF07_BC, CRF08_BC, and B by this assay, giving a sensitivity of 91.7% (66/72), and a specificity of 100%.
CONCLUSION
This assay is a simple, fast, and cost-effective subtyping method for HIV-1 CRF01-AE, CRF07_BC, CRF08_BC, and B strains prevailing in Guangxi.
目的
建立一种快速巢式多重PCR检测方法，用于对广西流行的HIV-1 CRF01_AE、CRF07_BC、CRF08_BC、B和C亚型毒株进行基因分型。
方法
根据这些亚型的gag序列设计亚型特异性引物。分别采用巢式多重PCR、DNA测序和系统发育分析对广西HIV-1样本的亚型进行鉴定，然后计算巢式多重PCR的敏感性和特异性。
结果
巢式多重PCR能够正确鉴定5份已知亚型样本，对所有HIV阴性样本均无反应。在72份HIV阳性样本中，该检测方法正确鉴定出66份为CRF01_AE、CRF07_BC、CRF08_BC和B亚型，敏感性为91.7%（66/72），特异性为100%。
结论
该检测方法是一种用于广西流行的HIV-1 CRF01-AE、CRF07_BC、CRF08_BC和B亚型毒株基因分型的简单、快速且经济高效的方法。
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