本文選題：HIV-1　切入點：嬰兒　出處：《中國疾病預防控制中心》2013年碩士論文　論文類型：學位論文

【摘要】：研究背景及目的 在中國隨著艾滋病疫情由高危人群向一般人群擴散,性傳播已經逐漸成為HIV(Human Immunodeficiency Virus)主要傳播途徑,女性感染者比例增加,從而使預防艾滋病母嬰傳播(MTCT, Mother-to-child Transmission)面臨很大的挑戰(zhàn)。目前歐美發(fā)達國家主要使用高效抗反轉錄病毒治療策略(HAART, Highly Active Antiretroviral Treatment)預防艾滋病母嬰傳播,使得艾滋病母嬰傳播率控制在2%以下�？共《舅幬锏氖褂秒m然降低了艾滋病母嬰傳播率,但也存在產生耐藥突變的風險。在1998-1999年及2001-2002年美國紐約的回顧性調查研究中發(fā)現(xiàn),HIV-1感染嬰兒中耐藥突變率分別為12.1%和19%。目前中國艾滋病嬰兒的耐藥情況還沒有比較詳細的研究。在本研究中,我們使用干血斑樣本(DBS, Dried Blood Spots)對中國部分地區(qū)HIV-1感染嬰兒進行基因型耐藥檢測,同時分析嬰兒HIV-1亞型分布及V3環(huán)氨基酸特征。 研究方法 1.使用In-house方法進行基因型耐藥檢測； 2.巢式PCR擴增HIV-1env區(qū)結合pol進行基因分型并對env區(qū)V3環(huán)氨基酸進行分析。 研究結果 1.共檢測65名嬰兒的100份HIV-1DNA陽性DBS樣本,其中57名嬰兒的84份DBS樣本Pol區(qū)擴增成功,耐藥情況為：非核苷類反轉錄酶抑制劑;NNRTIs)耐藥突變占15.79%(9of57)；核苷類反轉錄酶抑制劑;(NRTIs)耐藥突變占8.77%(5/57)；沒有發(fā)現(xiàn)蛋白酶抑制劑主要突變。 2.共檢測65名嬰兒的100份HIV-1DNA陽性DBS樣本,其中45名嬰兒pol和env區(qū)均擴增成功,根據(jù)兩基因區(qū)進行分型：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區(qū)擴增成功的63份DBS樣本中V3環(huán)頂端四肽存在五種類型：GPGQ、 GPGR、GPGS、GLGR、GQGR,其中以GPGQ為主。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.對env區(qū)測序成功的63份DBS樣本輔助受體預測：CCR5輔助受體占87.30%(55/63),CCR4輔助受體占4.76%(3/63),未能做出預測占7.94(5／63)。CRF_BC亞型中42份使用CCR5受體,3份樣本未能做出預測；CRF_01AE亞型中,1l份使用CCR5輔助受體,2份樣本未能做出預測；B’亞型中3份使用CXCR4受體；G亞型中2份樣本使用CCR5輔助受體。 研究結論 1.在我國HIV-1感染嬰兒中,已經出現(xiàn)抗病毒藥物耐藥相關突變,建議HIV-1陽性孕婦在孕期服用抗病毒藥物前或用藥一段時間后進行耐藥檢測,根據(jù)耐藥檢測結果選取適宜的母嬰傳播阻斷藥物。 2.中國部分地區(qū)HIV-1感染嬰兒病毒亞型以CRFBC重組和CRF_01AE亞型為主。 3.嬰兒HIV-1env區(qū)V3環(huán)頂端四肽氨基酸分布存在多態(tài)性,包括GPGR、GPGQ、 GQGR、GPGS、GLGR五種類型,其中CRFBC、CRF_01AE亞型主要以GPGQ為主,在診斷試劑、輔助受體拮抗劑及疫苗的研發(fā)中應予以考慮。 4.嬰兒樣本中,預測HIV-1進入CD4細胞輔助受體主要為CCR5輔助受體,提示后續(xù)用藥可以考慮使用CCR5拮抗劑。
[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.
【學位授予單位】：中國疾病預防控制中心
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R725.1;R440

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