结核性脑膜炎临床分离株的耐药性和基因型相关性分析

目的 了解结核性脑膜炎（tuberculous meningitis,TBM）的致病菌在耐药性和基因型方面的相关性.方法 从500例疑似TBM患者脑脊液标本中经MGIT（Mycobacteria growth indicator tube）960培养仪分离培养出25株结核分枝杆菌（Mycobacterium tuberculosis,Mtb）,对这25株Mtb分别进行间隔区寡核苷酸分型（Spoligotyping）法基因分型、14种抗结核药物的药物敏感性试验、10个耐药基因测序.结果 20株为北京基因型,占80.0％（20/25）;耐多药结核性脑膜炎（MDR TBM）有3株（3/25）,都属于北京基因型;测序结果显示inhA、embB、gyrB、eis 基因未发生突变,katG、rpoB、gyrA、rrs-KAN、rpsL、gidB基因发生突变.结论 北京基因型在耐药TBM中所占比例高,尤其是MDR-TBM.耐药TBM临床分离株与相关耐药基因突变有关系.     

深圳地区未经治疗艾滋病患者蛋白酶和逆转录酶耐药基因变异检测

目的 了解广东省深圳地区部分未经治疗的AIDS患者蛋白酶和逆转录酶耐药基因变异的发生频率和类型。方法 收集深圳市45例未经治疗的AIDS患者血浆，用RT-PCR及套式PCR扩增HIV-1pol区蛋白酶基因全序列与逆转录酶基因部分序列，将反应产物直接进行序列测定，并分析耐药基因的突变情况。结果 从45份血浆标本中扩增出41份pol区基因并进行序列测定与分析，蛋白酶基因耐药的主要突变发生率为12．2％（5／41），次要突变发生率为100％（41／41）；逆转录酶基因的耐药基因突变发生率为39．0％（16／41）。对上述耐药基因突变的评分表明，对蛋白酶类药物的耐药率为9．8％（4／41）；对逆转录酶类药物的耐药率为19．5％（8／41）；有1份标本对蛋白酶抑制剂、2份标本对逆转录酶抑制剂潜在耐药。结论 深圳地区未经治疗的AIDS患者中存在HIV-1耐药株，但多数患者对现有抗病毒药物敏感。应用抗病毒治疗必须加强监测，保持较好的依从性，防止耐药性毒株的流行。     

未经治疗的慢性乙型肝炎患者乙型肝炎病毒耐药变异、基因型和血清型研究

目的 研究未经核苷(酸)类似物(NA)治疗的慢性乙型肝炎患者HBV耐药变异、基因型、基因亚型和血清型特点.方法 从北京大学附属医院收集97例未经NA治疗的慢性乙型肝炎患者血清,用半巢式聚合酶链反应-直接测序法获得HBV全长逆转录酶区序列,用生物信息学技术筛查该区内11个经典耐药变异位点并鉴定基因型、基因亚型和血清型.用统计分析软件SPSS11.0进行t检验和χ~2检验. 结果 HBV在11个经典耐药变异位点上均为野生型氨基酸;B基因型和C基因型分别占36.1%(35/97)和63.9%(62/97),前者均属B2亚型,后者C2亚型占91.9%(57/62),C1亚型占6.5%(4/62),1例未能分出亚型.已知出生地的患者中,71.9%(23/32) B基因型感染者出生于我国南方地区,81.6%(40/49) C基因型感染者出生于北方地区,基因型地域分布特点明显,χ~2=23.19,P<0.01.血清型为adr者占60.8%(59/97),与C基因型相关;为adw者占38.1%(37/97),与B基因型相关,χ~2=87.83,P<0.01.结论 未经NA治疗的慢性乙型肝炎患者体内野毒株为优势株,其基因型、基因亚型和血清型与患者出生地有关.     

乙型肝炎病毒感染对艾滋病患者联合抗反转录病毒治疗效果的影响

目的 了解HBV感染对AIDS患者联合抗反转录病毒治疗(cART)效果的影响.方法 对HIV、HBV合并感染者78例和AIDS患者156例定期进行CD4+T淋巴细胞、HIV RNA、HBV血清学标志物和肝功能检测,并记录其生存情况,比较两组患者cART期间免疫学和病毒学应答的差异.计数资料采用卡方检验,计量资料采用t检验,非正态分布的计量资料采用两独立样本非参数检验.结果 cART第42个月时,同一治疗时间的CD4+T淋巴细胞和HIV RNA水平在HIV、HBV合并感染者与单纯AIDS患者间比较,差异均无统计学意义;cART第48、54和60个月时,HIV、HBV合并感染者免疫学和病毒学应答水平均低于单纯AIDS患者.HIV、HBV合并感染者在cART后第12、24、36、48和60个月时,13例患者中有3例在各时间点均表现为HBeAg阴转;抗-HBe阳转率分别为32.1％(9/28)、50.0％(14/28)、53.6％(15/28)、64.3％(18/28)和71.4％(20/28),阳转率逐年增高(x2=10.189,P=0.037); HBV DNA阴转率分别为95.1％(39/41)、82.9％(34/41)、68.3％(28/41)、43.9％(18/41)和43.9％(18/41),阴转率逐年下降(x2=29.982,P=0.000);肝功能异常率分别为32.1％(25/78)、51.4％(38/74)、33.8％(22/65)、47.9％(23/48)及6.7％(3/45),各时间点差异有统计学意义(x2=28.053,P=0.000).HIV、HBV合并感染者及单纯AIDS患者的病死率分别为24.4％(19/78)和5.1％(8/156),差异有统计学意义(x2=18.841,P＜0.01),且HIV、HBV合并感染者84.2％死于HBV相关的终末期肝病.结论 合并HBV感染可影响cART的远期疗效,终末期肝病是HIV、HBV合并感染者接受cART后的首要死因.     

艾滋病抗病毒治疗失败患者的耐药情况分析

目的:了解艾滋病患者抗病毒治疗失败后的耐药状况。方法:纳入2017年10月至2018年12月于郑州市第六人民医院行抗病毒治疗失败的156例艾滋病患者。人类免疫缺陷病毒(human immunodeficiency virus,HIV)-1耐药采用HIV-1 ViroSeq?基因型检测法,与斯坦福大学HIV耐药数据库(h...     

123株人类免疫缺陷病毒-1重组亚型主要耐药性突变的基因屏障

目的 比较HIV-1 CRF01_AE、CRF07_BC和CRF08_BC毒株发生蛋白酶抑制剂(PI)、核苷类反转录酶抑制剂(NRTI)和非核苷类反转录酶抑制剂(NNRTI)主要耐药性突变的基因屏障,分析不同亚型耐药模式的差异.方法 纳入在广西壮族自治区南宁、柳州两市未经治疗的HIV感染者190例,采集外周静脉血,从血浆中提取HIV-1 RNA,扩增pol区并测序,用系统进化树分析序列的亚型,统计各序列每个主要突变位点由野生型密码子突变为耐药密码子所需的碱基转换和颠换的数量,根据转换与颠换发生概率约2.5:1的规律,将每个转换赋值设为1,每个颠换赋值设为2.5,计算各突变赋值之和,即为基因屏障值,采用Kruskal-Wallis检验法和Nemenyi法比较不同亚型的基因屏障差异.结果 共获得CRF01_AE、CRF07_BC和CRF08_BC毒株123株.CRF08_BC发生T/S69D的基因屏障低于CRF01_AE和CRF07_BC(χ2=107.501,P＜0.01),CRF01_AE和CRF07_BC发生V118I和L210W的基因屏障低于CRF08_BC,CRF07_BC发生V106M的基因屏障低于CRF01_AE和CRF08_BC.结论 在相同的选择压力下,CRF01_AE和CRF07_BC比CRF08_BC易发生V118I和L210W,CRF08_BC比CRF01_AE和CRF07_BC易发生T/S69D,CRF07_BC比CRF08_BC和CRF01_AE易发生V106M.     

中国部分地区应用高效抗逆转录病毒治疗HIV-1患者的耐药性检测

目的了解我国应用高效抗逆转录病毒治疗的人类免疫缺陷病毒（HIV）感染者中耐药情况.方法收集我国上海、河南及安徽等地116份应用抗病毒治疗患者的血浆标本,采用逆转录聚合酶链反应扩增pol基因片段并进行序列测定,通过HIV耐药性数据库进行耐药相关性突变分析,并构建进化树.结果共获得44例HIV-1分离株的pol基因序列,有70%（31/44）的标本发现了针对常用的三类抗逆转录病毒药物的主要耐药性突变,其中有34%（15/44）的标本发现了针对核苷类逆转录酶抑制剂（NRTIs）的耐药性突变;有68%（30/44）的标本发现了针对非核苷类逆转录酶抑制剂（NNRTIs）的耐药性突变;仅有2%（1/44）的标本发现了对蛋白酶抑制剂（PIs）类药物的主要耐药性突变.亚型分析表明91%（40/44）为B亚型.结论在我国应用抗逆转录病毒治疗的患者中耐药性的产生已经十分严重,尤其是针对NNRTIs的高度耐药性较为突出,迫切需要应用耐药性检测进行指导.     

乙型肝炎病毒合并人类免疫缺陷病毒感染对抗反转录病毒治疗效果的影响

目的:了解乙型肝炎病毒(hepatitis B virus,HBV)合并人类免疫缺陷病毒(human immunodeficiency virus,HIV)感染对抗反转录病毒治疗(anti-retroviral therapy, ART)效果的影响。方法:纳入2016年9月至2019年10月重庆市公共卫生医疗救治中心收...     

获得性免疫缺陷综合征患儿抗病毒治疗后耐药基因变异分析

目的 观察接受联合抗病毒治疗获得性免疫缺陷综合征(AIDS)患儿人类免疫缺陷病毒(HIV)的蛋白酶、逆转录酶及蛋白酶切割位点的耐药基因变异。方法 利用套式逆转录聚合酶链反应(RT—PCR)从7例接受蛋白酶和逆转录酶抑制剂抗病毒治疗的AIDS患儿的血浆中扩增HIV的蛋白酶、逆转录酶、gag和部分pol基因，直接将PCR产物进行核苷酸序列测定或克隆后测序，分析测序结果，确定耐药基因变异。结果 有6例患者发生了多位点蛋白酶和逆转录酶耐药基因变异，其中2例患者在发生多位点蛋白酶和逆转录酶耐药基因变异的基础上出现了gag蛋白酶切割位点的耐药基因变异。结论 大部分长时间(33～106个月)接受蛋白酶和逆转录酶抑制剂联合抗病毒治疗的AIDS患儿体内HIV发生了蛋白酶和逆转录酶耐药基因变异，其中只有少数患儿的HIV发生了gag蛋白酶切割位点的耐药基因变异。     

HIV-1非核苷类反转录酶抑制剂耐药相关突变及其复制适应性研究

非核苷类反转录酶抑制剂是高效抗反转录病毒疗法(HAART)治疗艾滋病的重要组成部分。文章综述了非核苷类反转录酶抑制剂(NNRTIs)的耐药相关突变位点,如主要耐药突变位点、罕见的耐药突变位点、非多态性附属耐药突变位点及突变位点的相互作用;并分别阐述了各个主要耐药突变位点对病毒复制适应性的影响。对今后HIV-1NNRTIs耐药相关工作进行了展望。     

Histoplasmosis in patients with the acquired immune deficiency syndrome

Five patients with disseminated histoplasmosis are reviewed. Four of five had the acquired immune deficiency syndrome (AIDS) and one was receiving steroid therapy. All were immigrants to the United States from Puerto Rico, the Dominican Republic, or South America, and none had a history of travel to regions of the United States where Histoplasma is endemic. Histoplasma complement fixation titers to mycelial antigen were not demonstrable in three of three patients in whom they were measured. Of the four patients with AIDS, Histoplasma capsulatum was isolated from bone marrow aspirates in two patients and from lymph node and liver biopsy specimens in one patient each. One of the bone marrow specimens showed organisms on Gomori-methenamine silver stain. In the other three cases, results of staining were falsely negative and diagnosis awaited culture results weeks later. Amphotericin B therapy resulted in rapid clinical improvement in the three patients that were treated. Intravenous therapy was followed by treatment with oral ketoconazole. Follow-up has not been long enough to determine the ultimate efficacy of ketoconazole. Disseminated histoplasmosis should be considered in all patients from the Caribbean or South America with AIDS or who are receiving immunosuppressive therapy.     

Cryptococcal infections in patients with acquired immune deficiency syndrome

Cryptococcus neoformans is a major pathogen in patients with acquired immune deficiency syndrome and was found to infect 13.3 percent of such patients seen at two medical centers. Serum cryptococcal antigen levels were as high as 1:2,000,000 and, despite therapy, often remained elevated. Antigen titers in the cerebrospinal fluid generally declined at an expected rate in the survivors. The significance of high antigen titers in the blood after a prolonged course of therapy with amphotericin B and 5-flucytosine is unknown.     

Disseminated histoplasmosis in patients with the acquired immune deficiency syndrome

The ongoing epidemic of acquired immune deficiency syndrome (AIDS) has affected homosexual men, intravenous (IV) drug abusers, Haitians, hemophiliacs, and others. Defects in cell-mediated immunity place these patients at risk for opportunistic infections. We recently saw three men from Alabama with disseminated infection due to Histoplasma capsulatum. Two of these men were homosexual and the other was an IV drug abuser. These three patients had evidence of depressed cellular immunity consistent with a diagnosis of AIDS. Infection caused by organisms indigenous to certain geographic areas of the United States may become more common in patients with AIDS as the epidemic continues.     

Helicobacter pylori infection in patients with acquired immune deficiency syndrome

A controlled study was conducted on patients with human immunodeficiency virus (HIV) infection referred for upper endoscopy to evaluate the prevalence of Helicobacter pylori (H. pylori) infection. Four different stains and culture for H. pylori were performed on biopsy specimens from the gastric antrum. Sixteen (40%) of 40 patients with acquired immune deficiency syndrome (AIDS) or AIDS-related complex (ARC) were diagnosed to be infected with H. pylori versus 14 (39%) of 36 age-matched control patients. Eight of 15 AIDS/ARC patients without AIDS-related esophagogastroduodenal findings (53%) were infected with H. pylori versus 8/25 (32%) with endoscopic findings typical of AIDS. No invasion of the lamina propria by H. pylori was noted in any patient. Active chronic gastritis was present in 60% of AIDS/ARC patients and 61% of controls. Fifty-eight and 59%, respectively, of active chronic gastritis cases were infected with H. pylori. All the H. pylori infections, except one, were found in patients with chronic gastritis. In AIDS/ARC patients, H. pylori infection and active chronic gastritis are as common as in other patients referred for upper endoscopy. They may play a pathogenic role, especially when endoscopic AIDS-related findings are lacking. Cell-mediated immune deficiency does not appear to increase the risk of infection with H. pylori.