越南东北部四省HIV-1亚型分布及其耐药性研究

发布时间：2018-02-10 07:50

本文关键词： 越南 HIV流行亚型耐药突变　出处：《广西医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的通过获得越南东北部四省接受抗病毒治疗人群的HIV-1 pol区序列,以期了解HIV毒株在越南北部的亚型分布和耐药特征。方法通过方便抽样,在越南北部广宁省,谅山省,高平省和何江省HIV-1流行严重地区选取接受抗病毒治疗的HIV感染者或患者作为研究对象。对研究对象进行流行病学调查,采集外周静脉血液10ml,提取血细胞前病毒DNA,采用巢式PCR对HIV-1 pol基因片段进行扩增,将阳性扩增产物送北京博迈德公司进行Sanger法测序,然后采用Bioedit、Chromas和MEGA等相关生物学软件对所得序列进行序列的比对校正、亚型分析。最后采用Spss 16.0软件对所得数据进行统计分析。结果 1.本次研究共招募433例感染者,采集自越南北部的4个省份,包括高平(112例)、河江(121例)、谅山(96例)、广宁(104例),分别占25.87%、27.94%、22.17%、24.02%,纳入研究的HIV-1感染者均接受高效抗逆转录病毒治疗;其中男性247例,占57.04%,男女比例为1.33:1;平均年龄为(38.52±8.18)岁,以25-50岁居多;婚姻状况以已婚、离异或丧偶、未婚为主,分别占61.66%、19.63%和14.55%;文化程度以初中以上学历为主,占88.91%;传播途径以注射吸毒为主,占52.66%,其次为异性性传播,占26.33%。2.本研究共收集433例HIV-1感染者的血浆样本,成功扩增出206例感染者的HIV-1 pol基因序列,测序进行基因分型,系统进化树显示206例样本均为CRF01_AE亚型。3.对获得的206例CRF01_AE亚型HIV-1毒株进行基因型耐药性分析,结果显示有32例样本至少对一种抗病毒药物,其中19例样本同时对NRTIs和NNRTIs类药物耐药,总耐药率为15.53%(32/206)。在32例耐药样本中,注射吸毒途径为13例(13.40%),异性性传播为8例(12.31%)。4.耐药率在不同省间的分布有差异,耐药发生率最高的是谅山省和广宁省,均占23.08%,其次是河江省(16.39%)、高平省(5.97%)。5.本次研究获得32例耐药毒株,其中NNRTIs和NRTIs双重耐药毒株最多,为19例,占59.38%;其次分别为NNRTIs(8例)、NRTIs(3例)和PIs(2例),分别占25.00%、9.38%和6.25%。6.在NNRTIs耐药的样本中,以181、190、230、179位置突变为主,其中突变位点Y181C、G190A、M230I和V179D分别占所有突变位点总数的19.40%、13.43%、13.43%和10.45%,共56.71%,占比超过其他突变位点总和(43.29%);其他位置如101、103、225等有少量分布。在NRTIs耐药的样本中,以184、75、215、41、65位置突变为主,其中突变位点M184V、V75M、T215F、M41L、K65R分别占所有突变位点总数的14.29%、10.00%、10.00%、8.57%、7.14%。在PIs耐药的样本中,以突变位点G73S(55.56%)居多,其他为I84V、K43T、L89V、L90M。7.统计学结果分析显示,NNRTIs不同药物的耐药程度差异有统计学意义(χ2=37.248,P0.001),NNRTIs耐药以中、高度耐药为主,占到总数的81.6%;中度耐药以ETR(26.19%)、RPV(38.10%)居多,高度耐药以EFV(28.57%)、NVP(42.86%)居多。NRTIs不同药物的耐药程度差异有统计学意义(χ2=39.445,P0.001),NRTIs耐药以高度耐药为主,约占到总数三分之二;高度耐药以FTC(29.57%)、3TC(29.51%)居多,其次是DDI(13.11%)、ABC(9.84%)、D4T(9.84%)、AZT(4.92%)、TDF(3.28%)。PIs不同药物的耐药程度差异无统计学意义(χ2=3.773,P=1.000)。影响耐药发生的logistics回归多因素分析,结果表明,样本来源于谅山和广宁省是基因型耐药发生的危险因素(P0.05)。结论:越南东北部四省的流行的HIV毒株以CRF01_AE亚型为主;东北部四省的抗病毒治疗人群HIV发生耐药突变率高于全越南其他地区,主要对NNRTIs和NRTIs药物耐药为主。
[Abstract]:Objective to sequence HIV-1 pol District in northeastern Vietnam four provinces receive antiviral treatment groups, in order to understand the HIV strains in northern Vietnam subtype distribution and drug resistance characteristics. By convenient sampling, Guangning Province in northern Vietnam, Lang Son Province, Gaoping province and Zhejiang Province HIV-1 epidemic area were receiving antiretroviral treatment of HIV infected persons or patients as the research object. Epidemiological survey was conducted on the research object, collected peripheral venous blood 10ml extracted from blood cells before virus DNA by nested PCR for HIV-1 pol gene fragment was amplified, the positive PCR products were sent to Beijing Bomaide company Sanger sequencing, and then using Bioedit, Chromas and MEGA and other related correction ratio biological software sequence in the sequence and subtype analysis. Finally, using Spss 16 statistical software to analysis the data. The results of this study to recruit a total of 1. Raised 433 cases of infection, 4 provinces gathered from more North and south, including Gaoping (112 cases), river (121 cases), Lang Son (96 cases), Guangning (104 cases), respectively 25.87%, 27.94%, 22.17%, 24.02%, HIV-1 infection were included in the study on HAART treatment; 247 cases were male, accounting for 57.04%, the ratio of male to female was 1.33:1; the average age was (38.52 + 8.18) years old, with 25-50 years of age; marital status to married, divorced or widowed, unmarried accounted for 61.66%, 19.63% and 14.55%; education in junior high school or above is the main route of transmission, accounting for 88.91%; mainly by injection drug use accounted for 52.66%, followed by heterosexual transmission accounted for 26.33%.2., plasma samples were collected for the study of 433 cases of HIV-1 infection, and successfully amplified the HIV-1 sequence of pol gene in 206 cases of infection, sequencing, genotyping, phylogenetic analysis showed that 206 cases were CRF01_AE subtype samples for.3. 206 cases of C RF01_AE subtype HIV-1 strains were genotype drug resistance analysis, results showed that 32 samples of at least one of antiviral drugs, including 19 cases of samples at the same time on NRTIs and NNRTIs drug resistance, total drug resistance rate was 15.53% (32/206). In 32 cases of drug samples, drug injection way for 13 cases (13.40%), specificity the spread of 8 cases (12.31%).4. resistance rate differences in the distribution of different province, the resistance rate is the highest in Lang Son province and Guangning Province, accounted for 23.08%, followed by the river province (16.39%), Gaoping province (5.97%).5. in this study were obtained from 32 patients with drug-resistant strains, including NNRTIs and NRTIs double resistant strains up to 19 cases, accounting for 59.38%; followed by NNRTIs (8 cases), NRTIs (3 cases) and PIs (2 cases), respectively 25%, 9.38% and 6.25%.6. in NNRTIs resistant samples, with 181190230179 point mutations, including mutations of Y181C, G190A, M230I and V179D respectively. For all Mutations in 19.40% of the total, 13.43%, 13.43% and 10.45%, a total of 56.71%, accounting for more than the sum of the other mutations (43.29%); other positions such as 101103225. A small amount of distribution in NRTIs resistant samples, 184,75215,41,65 mutation position, the mutation sites of M184V, V75M, T215F, M41L, K65R respectively in all mutant the total number of sites 14.29%, 10%, 10%, 8.57%, 7.14%. in PIs resistant samples, the mutation sites of G73S (55.56%) are the other for I84V, K43T, L89V, L90M.7. statistical analysis indicated that there were statistically significant differences in degree of resistance to NNRTIs of different drugs (2=37.248, P0.001), NNRTIs resistance to, highly resistant, accounting for 81.6% of the total; moderate resistance to ETR (26.19%), RPV (38.10%) are highly resistant to EFV (28.57%), NVP (42.86%).NRTIs are not the same degree of drug resistance was statistically significant (2=39.445, P0.001), The resistance of NRTIs to highly resistant, accounting for about 2/3 of the total number of highly resistant to; FTC (29.57%), 3TC (29.51%) majority, followed by DDI (13.11%), ABC (9.84%), D4T (9.84%), AZT (4.92%), TDF (3.28%) there was no significant difference in different degree of resistance to drugs (.PIs x 2=3.773, P=1.000). Impact analysis, multiple regression results show that the resistance of logistics, samples from Lang Son and Guangning province are risk factors for the occurrence of resistant genotypes (P0.05). Conclusion: the northeastern province of Vietnam four epidemic HIV strains to CRF01_AE subtype; four northeastern Province antiviral treatment population the occurrence of HIV resistance mutation rate was higher than that in other areas throughout the country, mainly on the NNRTIs and NRTIs drug resistance.

【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R512.91;R181.3

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