海南省抗病毒治疗失败HIV/AIDS患者HIV-1亚型及基因型耐药分析

目的 了解海南省经艾滋病抗病毒治疗(antiretroviral therapy,ART)失败HIV/AIDS患者的HIV-1基因型耐药特点,为海南省HIV/AIDS患者有效抗病毒治疗提供基础依据.方法 采集2014-2019年ART治疗失败的患者血样进行基因型耐药检测,分析HIV-1毒株亚型与耐药突变特点.结果 17...     

四川地区HIV/AIDS患者基因亚型及传播性耐药现状研究

  目的  分析四川地区初治HIV/AIDS患者的HIV基因亚型分布特点和传播性耐药现状。  方法  纳入四川地区住院且无抗反转录病毒治疗（ART）药物暴露史的成年HIV/AIDS患者,采用In-house测序法进行HIV基因测序,系统进化树判定HIV基因亚型,美国斯坦福大学HIV耐药数据库在线比对耐药突变位点并确定是否存在耐药及耐药种类、耐药水平。  结果  共纳入符合标准的120例患者,采集120份血液样本。87.5%样本HIV基因扩增成功（105/120）,HIV基因亚型分布特点:CRF01_AE占比46.67%（49/105）,CRF07_BC占比39.05%（41/105）,其他占比14.28%（15/105）。不同性别、年龄、民族、感染途径、耐药、HIVRNA水平、CD4水平等基因亚型的差异均无统计学意义（P>0.05）。25份样本检测到耐药突变位点（20.83%）,20份样本潜在耐药（16.67%）,5份样本传播性耐药（4.17%）。24份样本对非核苷类反转录酶抑制剂（NNRTIs）耐药,V179D/E位点突变频率最高,1份样本对蛋白酶抑制剂耐药,突变位点M46I,未发现核苷类反转录酶抑制剂耐药及整合酶抑制剂耐药。  结论  四川地区初治住院成年HIV/AIDS患者HIV基因亚型分布特点以CRF01_AE和CRF07_BC为主,传播性耐药率较低,以NNRTIs耐药为主。进行HIV基线耐药检测对制定有效ART方案具有重要意义。     

HIV感染者高效抗逆转录病毒治疗失败后的耐药研究

日的：研究杭—HIV治疗失败患者体内HIVRNA和DNA的基因变异情况，探讨其耐药机理从而指导临床用药，并对现有耐药数据库进行补充。方法：对德国各尔大学艾滋病中心10例杭HIV耐药患者血浆中抽提HIVRNA和DNA，分别对其逆转录酶区(RT)和蛋白酶区(PI)进行PCR扩增，产物回收后进行测序，得到的序列与国际标准株HXB2CG和Standford大学耐药数据库比较，从而找出变异位点，并结合临床进行分析讨论。结果：我们发现了一些能引起对某些药物产生相应耐药的有意义的突变位点，并与临床结果一致。我们还发现在一些患者中存在的突变，在Standford大学耐药数据库中还未报道。结论：HAART治疗失败者存在不同程度的逆转录酶区和蛋白酶区变异，逆转录酶区V197I，蛋白酶区K20T，K20I等变异可能与耐药有关。     

四川省越西县和昭觉县HIV抗病毒治疗停药患者耐药调查

目的 了解四川凉山州HIV抗病毒治疗停药患者的耐药情况。方法 在四川省凉山州选择抗病毒治疗人数较多的越西县和昭觉县两个治疗点,对其2018年HIV抗病毒治疗停药患者按照抗病毒治疗号顺序进行横断面调查,收集调查对象社会人口学、高危行为以及抗病毒治疗情况等数据;采集全血分离血浆检测病毒载量,提取核酸和pol基因区扩增测序进行耐药检测。结果 调查273例患者,54.7%（140/273）患者的病毒载量≥1 000 拷贝/mL;成功获得206份序列进行耐药分析,16.0%（33/206）患者检出耐药。病毒载量≥1 000拷贝/mL的比例随着停药时间的延长而增大,而耐药的比例随着停药时间的延长而减小。昭觉县的耐药率高于越西县（aOR=2.97,95%CI=1.17~7.54,P=0.022）;未婚的患者耐药率比在婚或同居的患者高（aOR=2.58,95%CI=1.27~7.40,P=0.042）。停药时间 >24个月患者的耐药率比停药时间1~<24个月低（aOR=0.32,95%CI=0.14~0.76,P=0.001）。近一年内发生无保护性行为的比例为72.2%。结论 HIV抗病毒治疗停药患者耐药和无保护性行为比例均高,需对该人群加强宣传教育,减少HIV毒株尤其是耐药株的传播。     

抗病毒治疗失败的艾滋病患者HIV-1基因型耐药变异的研究

目的 了解高效抗逆转录病毒治疗(HAART)失败的艾滋病患者HIV-1基因型耐药变异的情况,为临床治疗提供实验室参考数据.方法 收集2008年1月至2009年12月在深圳市第三人民医院接受HAART治疗失败的41例艾滋病患者的血浆,采用RT-PCR和套式PCR从血浆中提取病毒RNA,扩增目的基因片段,并对其进行序列测定和分析.结果 共获得38例抗病毒治疗失败者的基因序列,其中有3条序列显示无变异位点,在其他出现耐药变异的序列中K103N、G190A、Y181C、K101P、M184V、D67N、K70R、T215Y及K219R为最常见的变异位点;100%(35/35)治疗失败患者出现了对非核苷类逆转录酶抑制剂NVP或EFV的高中度耐药,超过50%的患者出现了对临床上目前常用的核苷类逆转录酶抑制剂AZT、3TC、IMT及DDI的高中度耐药,仅发现极少数治疗失败患者对蛋白酶抑制剂的高中度耐药;D4T+DDI+NVP治疗方案是最常见出现耐药的治疗组合;病毒耐药变异往往发生在抗病毒治疗后的2～3年.结论 HIV耐药变异株的出现是艾滋病患者临床上抗病毒治疗失败的主要原因,目前引起耐药突变主要是针对NNRTIs或NRTIs药物耐药,而对Pis的耐药在临床上尚比较少见,建议更换治疗组合时多考虑选用含有增强作用的Pis,从而达到更好地病毒抑制效果.     

Glycaemic consequences of HIV and highly active antiretroviral therapy: a pilot study and review of literature

The metabolic consequences of HIV and AIDS are accentuated in the setting of highly active antiretroviral therapy. Peripheral lipodystrophy, central adiposity, hyperlipidaemia, insulin resistance and diabetes mellitus are frequent associations of protease inhibitor containing highly active antiretroviral therapy regimens. Ninety patients aged 25-50 years (males 52, females 38), seropositive for HIV 1 and 2 or both were selected to see the glycaemic profiles in asymptomatic early HIV disease with CD4 counts > 100/microl and to compare this with the glycaemic profile of (a) advanced HIV disease (CD4 counts < 200/microl), not on highly active antiretroviral therapy and (b) advanced HIV disease (CD4 counts < 200/microl), on uninterrupted non-protease inhibitor highly active antiretroviral therapy > 6 months. All the patients were grouped into 3: (1) Group A: CD4 counts > 500/microl (n=37), highly active antiretroviral therapy naive, (2) group B: CD4 counts < 200/microl (n=21), not on highly active antiretroviral therapy, and (3) group C: CD4 counts < 200/microl, receiving uninterrupted non-protease inhibitor based highly active antiretroviral therapy for > 6 months (n=32). The fasting blood glucose, glycosylated Hb (HbA1c) levels, were measured in all the patients in 3 groups and significance of difference between means was calculated among various groups. Body weight and waist-hip ratio were also measured. The results were analysed and compared with other studies.     

广西崇左市4个边境市县抗病毒治疗HIV感染者耐药情况及基因型

目的 了解广西崇左市4个边境市县（大新县、宁明县、凭祥市、龙州县）接受抗病毒治疗的艾滋病病毒（HIV）感染者的基因型耐药情况及其影响因素,为该地区艾滋病治疗人群的抗病毒治疗（ART）方案调整和优化提供科学依据。方法 纳入2018年1月—2018年9月371例来自崇左市4个边境市县年龄≥18岁的HIV感染者,收集研究对象抗病毒治疗信息和pol区基因序列进行基因亚型和耐药情况分析。结果 获得371例患者的HIV pol区序列,对抗病毒药物产生耐药有62例,总体耐药率16.7%,耐药患者基因亚型以CRF01_AE为主,占72.6%,其次是CRF08_BC,占22.6%;耐药类型以非核苷类药物为主,占75.8%,耐药突变位点以K103N为主,主要对EFV和NVP产生耐药。多因素Logistic回归分析结果显示,地区来源于龙州县（OR=3.392,95%CI：1.093~10.525）、基因亚型为08BC(OR=9.431,95%CI：3.296~26.985),更换过治疗方案（OR=2.662,95%CI：1.241~5.708）与检出基因型耐药密切相关。基因距离分析显示CRF01_AE>CRF08_BC>CRF07_BC;不同亚型中,耐药组基因距离均大于非耐药组。结论 广西边境部分地区HIV/AIDS 患者抗病毒治疗耐药率较高,且高度耐药占比较高,需加强合理用药、HIV耐药检测,提高患者服药依从性。     

Superior virological efficacy of ritonavir-boosted protease inhibitor regimens compared to single protease inhibitor therapy

Low dose-ritonavir boosted protease inhibitors are increasingly being used for the first-line antiretroviral treatment, though their virological efficacy has just poorly been compared to alternative antiretroviral therapies. Here, we retrospectively investigated the virological responses of 316 protease inhibitor-naive HIV patients receiving highly active antiretroviral treatment based on a single (n = 256) or a ritonavir-boosted protease inhibitor (n = 60), both in the background of two nucleoside analogues. - By intent-to-treat analysis, a complete initial virological response was achieved in 71.8% of all patients in the single protease inhibitor group (indinavir: 76%, ritonavir: 67.5%, nelfinavir: 70.6%) and in 88.3% (p = 0.008) of patients treated with a boosted protease inhibitor (saquinavir/r: 71.4%, indinavir/r: 92.1%, lopinavir/r: 86.6%). The multivariate risk analysis identified boosted PI treatment as an independent predictor of a complete virological response (OR = 2.8, p=0.02). Viral rebound after an initial complete virological response was observed in 28% and 17% (p = 0.06) of patients receiving a single or a dual protease inhibitor, respectively. The rate of durable viral suppression over 12 months was 44.5% and 56.7% (p = 0.09) in the respective study cohorts. Ritonavir-boosted protease inhibitors therefore seem to induce a superior virological response rate and a higher degree of sustained virological suppression.     

HIV associated arthritis: case report and review of the literature

HIV infection can be associated with different types of arthropathies which are often underdiagnosed. We present the case of a 52 year old HIV positive man on highly active antiretroviral therapy including indinavir who developed an acute painful oligoarthritis. We present this case on HIV associated arthritis and include a review on other HIV specific types of arthritis (acute symmetric arthritis and painful articular syndrome) which are assumed as entities exclusively apparent in HIV patients. The pathophysiology of arthritis in HIV infected patients is not yet completely understood but a direct role of the HIV on the initiation of synovitis is suspected in some of them. Additionally, there is evidence that antiretroviral drugs, in particular the protease inhibitor indinavir, can lead to arthritic complications as well.     

Rates of disease progression by baseline CD4 cell count and viral load after initiating triple-drug therapy.

CONTEXT: Current recommendations for initiation of antiretroviral therapy in patients infected with human immunodeficiency virus type 1 (HIV) are based on CD4 T-lymphocyte cell counts and plasma HIV RNA levels. The relative prognostic value of each marker following initiation of therapy has not been fully characterized. OBJECTIVE: To describe rates of disease progression to death and AIDS or death among patients starting triple-drug antiretroviral therapy, stratified by baseline CD4 cell count and HIV RNA levels. DESIGN, SETTING, AND PARTICIPANTS: Population-based analysis of 1219 antiretroviral therapy-naive HIV-positive men and women aged 18 years or older in British Columbia who initiated triple-drug therapy between August 1, 1996, and September 30, 1999. MAIN OUTCOME MEASURE: Cumulative mortality rates from the initiation of triple-drug antiretroviral therapy to September 30, 2000, determined using various CD4 cell and plasma HIV RNA thresholds. RESULTS: As of September 30, 2000, 82 patients had died of AIDS-related causes, for a crude AIDS-related mortality rate of 6.7%. The product limit estimate (SE) of the cumulative mortality rate at 12 months was 2.9% (0.5%). In univariate analyses, a prior diagnosis of acquired immunodeficiency syndrome (AIDS), CD4 cell count, use of protease inhibitors, and HIV RNA level were associated with mortality. There was no difference in mortality by age or sex. Only CD4 cell count remained statistically significant in the multivariate analysis. After controlling for AIDS, protease inhibitor use, and plasma HIV RNA level at baseline, patients with CD4 cell counts of less than 50/microL were 6.67 (95% confidence interval [CI], 3.61-12.34) times and those with counts of 50/microL to 199/microL were 3.41 (95% CI, 1.93-6.03) times more likely to die than those with counts of at least 200/microL. CONCLUSION: Our data demonstrate uniformly low rates of disease progression to death and AIDS or death among patients starting antiretroviral therapy with CD4 cell counts of at least 200/microL. In our study, disease progression to death and AIDS or death was clustered among patients starting therapy with CD4 cell counts less than 200/microL.     

Allele-specific real-time PCR testing for minor HIV-1 drug resistance mutations: assay preparation and application to reveal dynamic of mutations in vivo

Background It is very important for the clinical management to test for minor HIV-1 resistance mutations accurately and sensitively. The conventional genotypic assays of HIV drug resistance detection based on sequencing can only discriminate the mutations which present in more than 20%-30%. The aim of this study was to evaluate allele-specific real-time PCR (ASPCR) to detect the resistance-related mutations located at positions 103, 184 and 215.Methods We developed the allele-specific PCR assay, using the most common drug resistance mutations in Chinese AIDS patients, K103N, M184V/I, T215F/Y as a model system. The standards were constructed by cloning the wild-type and mutant DNA fragments into the T-vector. We designed specific primers to discriminate mutant templates in the real-time PCR using SYBR green as a fluorescence reporter. And then we evaluated the ASPCR assay and tested 140clinical samples using this method.Results The sensitivities of ASPCR assay were 0.04% for K103N, 0.30% for M1841, 0.40% for M184V, 0.03% for T215F and 0.02% for T215Y. The intra-assay and inter-assay coefficients of variation were less than 0.42. One hundred and forty plasma samples were tested by ASPCR and dynamic resistance curves of ten patients were obtained.Conclusions Drug resistance emerged half a year after the start of antiretroviral therapy. The mutation of T215Yemerged 1 to 1.5 years after starting treatment and then increased rapidly. The ASPCR assay we developed was a sensitive, accurate and rapid method to detect the minor HIV-1 variants and it can provide earlier and more drug-resistance information for HIV research and AIDS antiretroviral therapy.     

Allele-specific real-time PCR testing for minor HIV-1 drug resistance mutations: assay preparation and application to reveal dynamic of mutations in vivo

Background It is very important for the clinical management to test for minor HIV-1 resistance mutations accurately and sensitively. The conventional genotypic assays of HIV drug resistance detection based on sequencing can only discriminate the mutations which present in more than 20%-30%. The aim of this study was to evaluate allele-specific real-time PCR （ASPCR） to detect the resistance-related mutations located at positions 103, 184 and 215.Methods We developed the allele-specific PCR assay, using the most common drug resistance mutations in Chinese AIDS patients, K103N, M184V/I, T215F/Y as a model system. The standards were constructed by cloning the wild-type and mutant DNA fragments into the T-vector. We designed specific primers to discriminate mutant templates in the real-time PCR using SYBR green as a fluorescence reporter. And then we evaluated the ASPCR assay and tested 140clinical samples using this method.Results The sensitivities of ASPCR assay were 0.04% for K103N, 0.30% for M1841, 0.40% for M184V, 0.03% for T215F and 0.02% for T215Y. The intra-assay and inter-assay coefficients of variation were less than 0.42. One hundred and forty plasma samples were tested by ASPCR and dynamic resistance curves of ten patients were obtained.Conclusions Drug resistance emerged half a year after the start of antiretroviral therapy. The mutation of T215Yemerged 1 to 1.5 years after starting treatment and then increased rapidly. The ASPCR assay we developed was a sensitive, accurate and rapid method to detect the minor HIV-1 variants and it can provide earlier and more drug-resistance information for HIV research and AIDS antiretroviral therapy.     

贵州省艾滋病患者耐药基因突变研究

目的研究艾滋病(AIDS)患者耐药基因突变情况,为防止AIDS耐药传播及流行提供科学依据。方法采用NueliSens EasyQ荧光实时定量PCR系统对治疗1年以上的AIDS患者进行病毒载量检测,病毒载量>1 000 U/ml者采用Abbott公司ViroSeqTMHIV-1 Genotyping System V2.0系统进行扩增、测序和基因突变分析。结果 1 088例AIDS患者中病毒载量>1 000 U/ml者45例,获得POL区基因序列30例,且均有不同程度基因突变,仅7例产生耐药,总耐药率为0.64%(7/1 088);发现交叉耐药2例。7例耐药的AIDS患者均出现至少1种耐药相关基因突变位点,突变率为23.3%(7/30)。7例AIDS患者均对非核苷类逆转录酶抑制剂产生耐药基因突变,其耐药突变位点为K103N 4例、G190A 2例、V179D 1例、Y181C 1例、K101E 1例、M230L 1例,临床主要表现为对地拉韦定、依非韦仑及奈韦拉平高度耐药,其中有2例对核苷类逆转录酶抑制剂产生耐药基因突变,突变位点为M184V,临床上表现为对拉米夫定和恩曲他滨高度耐药。未见蛋白酶抑制剂耐药基因突变。结论贵州省AIDS患者的耐药情况目前仍处于较低水平且耐药形式较为单一;核苷类逆转录酶抑制剂及非核苷类逆转录酶抑制剂对贵州省AIDS患者的广泛治疗和使用仍然有效;及时监控和掌握AIDS患者的耐药基因突变情况,对于指导临床用药尤为重要。     

云南省HIV-1 CRF01_AE毒株基因型耐药研究

目的 了解云南省HIV-1 CRF01_AE毒株的流行分布及基因型耐药情况。 方法 收集2018—2021年云南省抗病毒治疗失败的HIV/AIDS患者的人口学资料,采集患者血浆,逆转录聚合酶链反应扩增HIV-1 pol基因蛋白酶和逆转录酶区并进行序列测定,对基因型为CRF01_AE的序列进行耐药情况分析。结果 通过基因型耐药检测共获得967例亚型为CRF01_AE序列,其中男性682例（占70.5%）,耐药509例,耐药率为52.6%,异性性传播813例（占84.1%）,同性性传播95例（占9.8%）。同性性传播的耐药率最高,为65.3%;治疗时长<12个月的耐药率最低,为35.9%,36~<48个月耐药率最高,为58.1%。核苷类反转录酶抑制剂（NRTIs）耐药率为33.9%,非核苷类反转录酶抑制剂（NNRTIs）的耐药率为48.5%,蛋白酶抑制剂（PIs）的耐药率较低为1.3%。NRTIs类药物中阿巴卡韦（ABC）、恩曲他滨（FTC）和拉米夫定（3TC）耐药率较高,分别为为32.9%、32.4%和32.4%;NNRTIs类药物中奈韦拉平（NVP）和依非韦仑（EFV）耐药率较高,分别为47.6%和47.5%,均以高度耐药为主。一线治疗方案耐药率56.2%高于二线治疗方案耐药率29.5%,差异有统计学意义（P<0.001）。NRTIs耐药突变位点中,以M184V/I突变位点为主,突变率为30.0%;NNRTIs耐药突变位点中,以K103N/Q/S突变位点为主,突变率为28.1%;PIs耐药位点突变率相对较低,主要的耐药突变位点是L33F,突变率为3.7%。结论 云南省抗病毒治疗失败亚型CRF01_AE的患者耐药率有所降低,但部分地区及人群的耐药率仍较高,应加强对重点地区和人群的管理,加强耐药监测,对治疗失败的患者及时掌握耐药情况,合理调整治疗方案。