云南省抗病毒治疗失败的人类免疫缺陷病毒感染/艾滋病患者中人类免疫缺陷病毒1型pol区多态性位点的分布

Distribution of polymorphic loci in human immunodeficiency virus type 1 pol region in human immunodeficiency virus infection/acquired immunodeficiency syndrome patients failing anti-retroviral therapy in Yunnan Province

目的探讨人类免疫缺陷病毒感染/艾滋病(HIV/AIDS)患者接受高效抗反转录病毒治疗(HAART)过程中耐药的发生与多态性位点分布规律间的关系。方法纳入2015年6月至2021年12月云南省16个地级行政区成功扩增得到pol区基因序列的HAART失败HIV/AIDS患者,采用人类免疫缺陷病毒(HIV)局部序列排比检索基本工具(BLAST)对序列进行亚型鉴定,MEGA 6.0软件进行亚型验证。将样本序列提交至美国斯坦福大学HIV耐药数据库进行耐药位点比对。分析不同耐药程度、治疗方案和HIV-1亚型患者的多态性位点分布情况。采用趋势χ^(2)检验分析不同耐药程度患者多态性位点出现频率变化趋势。统计学比较采用χ^(2)检验并作事后两两比较。结果1453例患者扩增获得序列,耐药检测结果为敏感者954例,潜在或低度耐药者224例,中度耐药者189例,高度耐药者86例。随着HIV-1耐药程度的增加,HIV-1蛋白酶区(PR区)I15V、L19I、D60E和反转录酶区(RT区)E36A、T39D、S48T位点出现频率呈下降趋势(χ^(2)趋势=19.86、9.16、13.66、37.64、18.44、40.86,均P<0.01),PR区V77I和RT区K122E呈上升趋势(χ^(2)趋势=12.19、10.03,均P<0.01)。齐多夫定+拉米夫定+洛匹那韦/利托那韦、齐多夫定+拉米夫定+依非韦伦、齐多夫定+拉米夫定+奈韦拉平和替诺福韦+拉米夫定+依非韦伦治疗组患者PR区E35D、M36I、D60E和RT区S48T、K122E、R211K位点的频率差异均有统计学意义(χ^(2)=22.46、9.32、14.46、26.85、18.92、24.26,均P<0.05)。经两两比较,E35D、M36I、D60E位点在齐多夫定+拉米夫定+洛匹那韦/利托那韦组的频率,S48T在齐多夫定+拉米夫定+依非韦伦组的频率,K122E在齐多夫定+拉米夫定+奈韦拉平组的频率,以及R211K在替诺福韦+拉米夫定+依非韦伦组的频率分别高于其他3组,差异均有统计学意义(均P<0.008)。流行重组型(CRF)08BC、CRF07BC和CRF01AE亚型患者中PR区T12S、I15V、L19I、M36I、V77I、L89M和RT区E53D、I135V、S162C、R211K、K277R位点的频率差异均有统计学意义(χ^(2)=693.60、712.51、798.11、434.85、386.91、657.78、932.58、409.21、344.39、469.44、260.48,均P<0.001)。经两两比较,T12S、I15V、L19I、E53D、I135V、S162C、R211K在CRF08BC亚型中的频率,V77I和K277R在CRF07BC亚型中的频率,以及M36I和L89M在CRF01AE亚型中的频率分别高于其他2组,差异均有统计学意义(均P<0.017)。结论HAART失败所产生的多态性位点在不同的耐药程度、治疗方案和HIV亚型等方面呈现出不同的分布特点,具有一定的特异性和共性,需加强对部分多态性位点的监测。

Objective To explore the relationship between drug resistance occurrence and the distribution pattern of polymorphic loci in individuals with human immunodeficiency virus infection/acquired immunodeficiency syndrome(HIV/AIDS)treated with highly active anti-retroviral therapy(HAART).Methods HAART-failed HIV/AIDS patients who successfully amplified the gene sequences of the pol region between June 2015 and December 2021 from 16 prefecture-level administrative regions in Yunnan Province were included.The resistant sequences were classified using the human immunodeficiency virus(HIV)basic local alignment search tool(BLAST)and validated through MEGA 6.0,and the obtained sequences were submitted to the Stanford University HIV Drug Resistance Database to identify drug resistance loci.The distribution of polymorphic loci was analyzed across patients exhibiting varying degrees of drug resistance,different treatment regimens and distinct HIV-1 subtypes.Changes of the frequencies of polymorphic loci in patients with different degrees of drug resistance were analyzed using trend chi-square test.Statistical comparisons and further paired comparisons were performed using chi-square test.Results Gene sequences were amplified from 1453 patients,and the resistance testing results showed 954 sensitive,224 potentially or low resistant,189 moderately resistant,and 86 highly resistant patients.The frequencies of mutations I15V,L19I,D60E in the HIV-1 protease region(PR region)and E36A,T39D,S48T mutations in the HIV-1 reverse transcriptase region(RT region)showed a decreasing trend as the degree of HIV-1 resistance escalated(χ^(2)trend=19.86,9.16,13.66,37.64,18.44 and 40.86,respectively,all P<0.01).Conversely,the mutations V77I in the PR region and K122E in the RT region showed an ascending trend(χ^(2)trend=12.19 and 10.03,respectively,both P<0.01).Distinct treatment groups,namely zidovudine(AZT)+lamivudine(3TC)+lopinavir/ritonavir(LPV/r),AZT+3TC+efavirenz(EFV),AZT+3TC+nevirapine(NVP),and tenofovir(TDF)+3TC+EFV,were examined.Statistically significant differences in the frequencies of mutations E35D,M36I,and D60E in the PR region,as well as S48T,K122E,and R211K in the RT region,were observed among these treatment groups(χ^(2)=22.46,9.32,14.46,26.85,18.92 and 24.26,respectively,all P<0.05).In paired comparisons,AZT+3TC+LPV/r group displayed higher frequencies of E35D,M36I,and D60E mutations,the AZT+3TC+EFV group showed a higher frequency of S48T mutation,the AZT+3TC+NVP group showed a higher frequency of K122E mutation,and the TDF+3TC+EFV group exhibited a higher frequency of R211K mutation,all with statistically significant differences(all P<0.008).The differences in the frequencies of T12S,I15V,L19I,M36I,V77I,L89M in the PR region and E53D,I135V,S162C,R211K,K277R in the RT region among circulating recombinant form(CRF)08_BC,CRF07_BC and CRF01_AE subtype group were statistically significant(χ^(2)=693.60,712.51,798.11,434.85,386.91,657.78,932.58,409.21,344.39,469.44 and 260.48,respectively,all P<0.001).In paired comparisons,the frequencies of T12S,I15V,L19I,E53D,I135V,S162C and R211K in CRF08_BC subtype,the frequencies of V77I and K277R in CRF07_BC subtype,and the frequencies of M36I and L89M in CRF01_AE subtype were higher than those in the other two groups,and the differences were all statistically significant(all P<0.017).Conclusions The polymorphic loci resulting from HIV-1 HAART failure show different distribution patterns across various degrees of drug resistance,treatment regimens and HIV-1 subtypes.These loci demonstrate both specific and shared characteristics.It is necessary to enhance the surveillance of select polymorphic loci.