摘要
目的:建立适用于艾滋病母婴传播早期诊断的HIV基因芯片检测技术。方法:在产后180d内,采集HIV抗体阳性母亲所生婴幼儿的样本进行检测。从HIV基因组Gag区选取3对适宜引物,经逆转录-聚合酶链反应(RT PCR)扩增3个HIV目的基因片断。扩增产物纯化后与载体连接,转化到JM10 9宿主菌,接种平板培养,提取质粒。将探针点于尼龙膜上,制成基因芯片。样本提取DNA后与阳性对照质粒混合,地高辛标记显色观察结果,并和抗体检测结果相比较。结果:采用HIV基因芯片对9位HIV抗体阳性母亲所生婴幼儿进行检测,其中阳性2名,阴性7名,与抗体确认结果相比,敏感性和特异性均为10 0 0 %。结论:该方法能够对婴幼儿进行早期诊断,判断HIV母婴传播的方式,便于干预和治疗。
Objective:To study the technology for establishing DNA microarrays for HIV-1 early diagnosis of mother to child transmission.Methods:The infants born from HIV-infective mothers were tested with DNA microarrays before 180 days. Three pairs of appropriate primers which were selected from Gag region of HIV-1 gene segments were amplified to three HIV-1 target segments by method of RT-PCR.The amplified production were connected with Vector after it had purified,translated to JM109 host bacteria,brought up in plain agar culture medium .The plasmids were extracted from positive bacterial colony. The probes were dipped on the nylon membrane to make DNA microarrays.The DNA which extracted from the sample were mixed with positive plasmids.The DNA microarrays were hybridized with Digoxigenin labeled probes ,then dyed to show the results compared with HIV-Ab.Results:9 infants were born from HIV-infected mothers were tested with DNA microarrays.Of them were 2 positive samples and 7 negative samples.Both sensitivity and the specificity was 100.0%.Conclusion:The DNA microarrays can be applied for early diagnosis of HIV-1 MTCT.It is good at intervention measure to the infants through estimate mode of HIV-1 MTCT.
出处
《现代预防医学》
CAS
北大核心
2005年第6期639-641,共3页
Modern Preventive Medicine
