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目的探索艾滋病病毒Ⅰ型(HIV-1)阳性孕产妇新生婴儿核酸检测的辅助性诊断方法,了解深圳地区母婴传播所感染的HIV-1阳性婴儿HIV-1毒株的分子流行情况,帮助分析HIV-1在该人群中传播的危险因素。方法收集深圳市2000-2007年19例HIV-1阳性孕产妇的血样,她们所生新生儿中28人的血样,应用套式聚合酶链式反应(Nested-PCR),对该样本膜蛋白基因(env基因)和核心蛋白(gag基因)进行扩增,并对其各基因区核苷酸序列进行测定和分析。结果28例婴儿中共得到6例PCR诊断为阳性的婴儿样本,6例样本中共存在CRF01-AE和CRF07-BC两种重组毒株以及B一种亚型,其在所有分析样本中的比例分别为4/6、1/6和1/6;在env基因区与对应的流行代表株01-AE.TH.90.CM240的基因离散率为(9.200±1.600)%;CRF01-AE重组株组内离散率为(11.700±4.000)%;在gag基因区与对应的流行代表株01-AE.TH.90.CM240、07-BC.CN.97.97CN001和B.CN.RL42的基因离散率分别为(4.075±0.763)%、(5.500±0.566)%和(7.150±1.485)%;CRF01-AE重组株、CRF07-BC重组株和B亚型组内离散率分别为(4.033±1.692)%、0.800%和1.900%。结论该方法有望成为HIV母婴垂直传播中新生婴儿的早期辅助性诊断方法。在深圳地区HIV-1阳性婴儿中,HIV-1流行株以CRF01-AE重组亚型为主,其次是B亚型。并且在母婴传播过程中,HIV-1env和gag基因变异性较小。
Objective To explore the diagnostic method of neonatal nucleic acid detection in HIV-1-positive pregnant women and to find out the molecular epidemic situation of HIV-1-positive infants infected by mother-to-infant transmission in Shenzhen area, Risk factors for HIV-1 transmission in this population. Methods The blood samples of 19 HIV-1-positive pregnant women from 2000 to 2007 in Shenzhen were collected and the blood samples of 28 newborn infants were collected. The nested PCR was used to detect the expression of membrane protein gene (env gene) and core protein (gag gene) were amplified and their nucleotide sequences were determined and analyzed. Results Six infants from 28 infants were diagnosed as positive by PCR. Two CRF01-AE and CRF07-BC recombinant strains and one subtype B were co-existed in 6 samples, and their proportions in all samples were 4/6, 1/6, and 1/6 respectively; the genetic divergence rate between the env gene region and the corresponding popular representative strain 01-AE.TH.90.CM240 was (9.200 ± 1.600)%; within the CRF01-AE recombinant strain group The discrete rate was (11.700 ± 4.000)%. The genetic divergence rates between the gag gene region and the corresponding popular representative strains 01-AE.TH.90.CM240, 07-BC.CN.97.97CN001 and B.CN.RL42 were (4.075 ± 0.763)%, (5.500 ± 0.566)% and (7.150 ± 1.485)%, respectively. The discrete rates of CRF01-AE recombinant strains, CRF07-BC recombinant strains and B subtypes were 4.033 ± 1.692% and 0.800 % And 1.900%. Conclusion This method is expected to be an early adjuvant diagnostic method for newborn infants with HIV-1 transmission in the mother-to-infant transmission. Among HIV-1-positive infants in Shenzhen, HIV-1 strains were predominantly CRF01-AE recombinant subtypes, followed by subtype B. And HIV-1env and gag genes have less variability during mother-to-child transmission.