中国部分地区HIV-1感染婴儿基因型耐药及亚型研究

发布时间：2018-03-20 14:41

本文选题：HIV-1　切入点：婴儿　出处：《中国疾病预防控制中心》2013年硕士论文　论文类型：学位论文

【摘要】：研究背景及目的在中国随着艾滋病疫情由高危人群向一般人群扩散,性传播已经逐渐成为HIV(Human Immunodeficiency Virus)主要传播途径,女性感染者比例增加,从而使预防艾滋病母婴传播(MTCT, Mother-to-child Transmission)面临很大的挑战。目前欧美发达国家主要使用高效抗反转录病毒治疗策略(HAART, Highly Active Antiretroviral Treatment)预防艾滋病母婴传播,使得艾滋病母婴传播率控制在2%以下。抗病毒药物的使用虽然降低了艾滋病母婴传播率,但也存在产生耐药突变的风险。在1998-1999年及2001-2002年美国纽约的回顾性调查研究中发现,HIV-1感染婴儿中耐药突变率分别为12.1%和19%。目前中国艾滋病婴儿的耐药情况还没有比较详细的研究。在本研究中,我们使用干血斑样本(DBS, Dried Blood Spots)对中国部分地区HIV-1感染婴儿进行基因型耐药检测,同时分析婴儿HIV-1亚型分布及V3环氨基酸特征。研究方法 1.使用In-house方法进行基因型耐药检测； 2.巢式PCR扩增HIV-1env区结合pol进行基因分型并对env区V3环氨基酸进行分析。研究结果 1.共检测65名婴儿的100份HIV-1DNA阳性DBS样本,其中57名婴儿的84份DBS样本Pol区扩增成功,耐药情况为：非核苷类反转录酶抑制剂;NNRTIs)耐药突变占15.79%(9of57)；核苷类反转录酶抑制剂;(NRTIs)耐药突变占8.77%(5/57)；没有发现蛋白酶抑制剂主要突变。 2.共检测65名婴儿的100份HIV-1DNA阳性DBS样本,其中45名婴儿pol和env区均扩增成功,根据两基因区进行分型：CRF_01AE占20.00%(9/45),CRF BC占73.33%(33/45),B’占4.44%(2/45),G占2.22%(1/45). 3.Env区扩增成功的63份DBS样本中V3环顶端四肽存在五种类型：GPGQ、 GPGR、GPGS、GLGR、GQGR,其中以GPGQ为主。GPGQ占90.48%(57/63)；GPGR占1.59%(1/63)；GPGS占3.17%(2/63)；GLGR占1.59%(1/63)；GQGR占3.17%(2/63)。 4.对env区测序成功的63份DBS样本辅助受体预测：CCR5辅助受体占87.30%(55/63),CCR4辅助受体占4.76%(3/63),未能做出预测占7.94(5／63)。CRF_BC亚型中42份使用CCR5受体,3份样本未能做出预测；CRF_01AE亚型中,1l份使用CCR5辅助受体,2份样本未能做出预测；B’亚型中3份使用CXCR4受体；G亚型中2份样本使用CCR5辅助受体。研究结论 1.在我国HIV-1感染婴儿中,已经出现抗病毒药物耐药相关突变,建议HIV-1阳性孕妇在孕期服用抗病毒药物前或用药一段时间后进行耐药检测,根据耐药检测结果选取适宜的母婴传播阻断药物。 2.中国部分地区HIV-1感染婴儿病毒亚型以CRFBC重组和CRF_01AE亚型为主。 3.婴儿HIV-1env区V3环顶端四肽氨基酸分布存在多态性,包括GPGR、GPGQ、 GQGR、GPGS、GLGR五种类型,其中CRFBC、CRF_01AE亚型主要以GPGQ为主,在诊断试剂、辅助受体拮抗剂及疫苗的研发中应予以考虑。 4.婴儿样本中,预测HIV-1进入CD4细胞辅助受体主要为CCR5辅助受体,提示后续用药可以考虑使用CCR5拮抗剂。
[Abstract]:Research background and purpose. With the spread of the AIDS epidemic from high-risk groups to the general population in China, sexual transmission has gradually become the main route of transmission of HIV(Human Immunodeficiency virus, and the proportion of women infected has increased. Thus, the prevention of mother-to-child transmission of HIV / AIDS (MTCT, Mother-to-child transmission) is facing a great challenge. At present, developed countries in Europe and the United States mainly use highly effective anti-retroviral treatment strategies (HAART, Highly Active Antiretroviral treatment) to prevent mother-to-child transmission of HIV. So that the mother-to-child transmission rate of AIDS is controlled below 2%. Although the use of antiviral drugs has reduced the rate of mother-to-child transmission of HIV, But there is also a risk of drug-resistant mutations. In a retrospective study conducted in New York in 1998-1999 and 2001-2002, it was found that the drug-resistant mutation rates of HIV-1 infected infants were 12.1% and 19 respectively. At present, the drug resistance of Chinese AIDS infants has not been compared with that of AIDS infants. In this study, We used Dried Blood samples to detect the genotype resistance of infants infected with HIV-1 in some regions of China. We also analyzed the distribution of HIV-1 subtypes and the characteristics of V3 cyclic amino acids in infants. Research method. 1. Genotypic drug resistance was detected by In-house method. 2. Nested PCR amplification of HIV-1env region combined with pol for genotyping and analysis of V3 cyclic amino acids in env region. Research results. 1. A total of 100 HIV-1DNA positive DBS samples were detected from 65 infants, of which 84 DBS samples from 57 infants were successfully amplified by Pol. The drug resistance of NNRTIs-NNRTIs15.79% and 8.7775% of 57%, respectively. No major mutations of protease inhibitors were found in NNRTIs-NNRTIs-resistant cases of NNRTIs-NNRTIs-NNRTIs-NNRTIs-NNRTIs-resistant mutants were 15.79%, 8.77% and 5.77%, respectively, and no major mutations in protease inhibitors were found. 2. A total of 100 samples of HIV-1DNA positive DBS were detected in 65 infants, of which 45 infants were successfully amplified in the pol and env regions. Two genotypes of CRF01AE accounted for 20.00% and 73.3333% 45% B' for 4.44%, 2.2245G% 2.2245G / 45% for CRF01AE and 20.00% for 73.3345% B', 4.44%, 2.2245G, 2.2245G, 2.2245G, 2.2245G, 2.2245G, 2.2245G, 2.2245G and 2.2245G, respectively. 3. In 63 successful DBS samples, there were five types: GPGQ, GPGRG, GPGRG, GPGQ, GPGQ, GPGQ, GPGR, GPGQ, GPGR, GPGQ, GPGQ, GPGR, GPGQ, GPGR, GPGR, GGGR, GGGR, GGGR, GPGQ, GGGR, GGGR, GGGR, GGGR, GGGR, GGGR, GGGR, GGGR, GGGR, GGGR, GGGR, GGGR, GGGR, GGGR, GGGR, GGGR, GGGR, GGGR, GQG@@. 4. Prediction of 63 DBS coreceptors successfully sequenced in env region; 87.30% of CCR5 coreceptors account for 4.76A / 63% of CCR4 coreceptors; 42 out of 7.94 / 5 / 63C. CRFBC subtypes have failed to predict the use of CCR5 receptors in 3 samples of CRF01AE subtypes. Two coreceptor samples failed to predict the use of CCR5 coreceptors in 3 of the B'subtypes using CXCR4 receptor G subtype. Research conclusion. 1. In Chinese infants infected with HIV-1, there have been mutations associated with antiviral drug resistance. It is suggested that pregnant women with positive HIV-1 should be tested for drug resistance before or after taking antiviral drugs during pregnancy. According to the results of drug resistance test, suitable drugs for blocking mother-to-child transmission were selected. 2. The major subtypes of HIV-1 infantile virus infection in China are CRFBC recombination and CRF_01AE subtype. 3. The distribution of tetrapeptide amino acids at the top of the V3 ring in infants with HIV-1env was polymorphic, including GPGRGQ and GQGRGR, among which GPGQ was the main subtype of CRFBCU CRF01AE, which should be considered in the research and development of diagnostic reagents, auxiliary receptor antagonists and vaccines. 4. In infant samples, it was predicted that the main coreceptor of HIV-1 entering CD4 cells was CCR5 coreceptor, suggesting that the use of CCR5 antagonist could be considered for subsequent drug use.
【学位授予单位】：中国疾病预防控制中心
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R725.1;R440

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