Time trends of paediatric HIV infection in North India

Paediatric HIV infection continues to pose a serious threat in the developing world. While in the developed world, mother to child transmission has been reduced to less than 3%, in India no regular zidovudine (azidothymidine) intervention programmes operate. Some 20 million babies are born each year and the number of infected babies could be >50,000 per year. The present study was designed to assess the change, if any, in the time trends of HIV infection in children over the last 15 years as observed at the surveillance centre attached to Nehru Hospital, Chandigarh. All patients reporting to the surveillance centre at the PGIME&R, Chandigarh were subjected to a detailed history and screened for HIV by the three tests protocol recommended by the WHO. In babies under 18 months of age, viral load assay or DNA analysis was done to confirm infection. Timetrends were ascertained over a 15-year period to assess the impact of information, education and communication programme launched by National AIDS Control Organisation. Data indicates that the total number of HIV positive cases increased 10-fold over the last 10 years. During 1991, 41 cases were recorded; the number increased to 439 in year 2001, and 574 in 2004 (r=0.98). A similar trend was observed in the paediatric age group. During the initial 5 years ie, 1987 to 1992 only 7 paediatric cases were documented positive while the number increased to 45 in the year 2001 to 64 in the year 2004 with a cumulative figure of 323 children. Linear regression analysis showed a highly significant trend (r=0.94). Out of the 323 cases, 44.6% were symptomatic. Maximum number of babies were observed in the age group of 3-5 years. Thirty-nine patients (12%) had acquired the infection through blood. Thus the information, education and communication programme has had very little impact on the HIV epidemic and it calls for urgent antiretroviral intervention in antenatal mothers to control the emerging paediatric HIV epidemic.     

昆明地区HIV和HCV共感染者丙型肝炎病毒基因型特征

目的分析昆明地区人类免疫缺陷病（HIV）和丙型肝炎病毒（HCV）共感染者丙型肝炎病毒基因型分布。方法对204例经逆转录聚合酶链反应检测HCV病毒栽量,核酸为阳性的样品,利用反向点杂交技术进行HCV基因分型检测,其中60例经免疫印迹试验（WB）确认的HIV感染者,对共感染和非共感染组的丙型肝炎病毒基因型分布进行分析。结果204例HCV患者中3b型85例（HCV感染率最高,为41．6％）;1b型HCV感染在HIV／HCV共感染组中的感染率（30．0％）明显高于单纯HCV感染组感染率（20．0％）（P〈0．05）。而3b型HCV感染在HIV／HCV共感染组中的感染率（35．0％）明显低于单纯HCV感染组感染率（44．2％）（P〈0．05）。1b型,3a型和3b型HCV合并HIV感染的发生率较高;2a型和6a型HCV合并HIV感染的发生率较低。不同丙肝基因型HIV／HCV共感染者间感染途径以静脉药隐为主,其次是性接触。结论昆明地区共感染纽中的HCV流行的基因亚型有1b,3a,3b,6a共4种,1h为流行的主要基因亚型3a,3b和6a型均占相当比例,初步说明昆明地区HCV流行的基因亚型呈现多样性．     

巢式多聚酶链反应快速检测胎儿性别

为建立一种非侵入性的对Ｘ性连锁遗传性疾病胎儿的早期产前性别鉴定方法,我们采用了巢式ＰＣＲ方法来扩增孕妇外周血中男性胎儿的Ｙ染色体特异序列,并与新生儿出生性别相对照。４４例孕妇中２２例分娩男性婴儿．其中１９例扩增出Ｙ特异片段,阳性检出率为８６．４％（１９／２２）．其余２２例分娩女性婴儿,其中无一例出现Ｙ特异扩增带,结果提示用巢式ＰＣＲ方法可快速判断胎儿性别,使临床上无伤性产前检查Ｘ性连锁遗传性疾病的胎儿性别成为可能。     

1型HIV感染多重PCR检测方法的建立及临床应用

目的建立检测人免疫缺陷病毒1型(HIV-1)的多重巢式PCR(nPCR)和多重反转录PCR(RT-PCR)方法。将其与已经上市的NASBA法试剂盒进行检测敏感性比较;探讨血浆RNA水平对PCR检测敏感性的影响;评价该方法用于我国HIV-1感染者诊断的价值。方法针对HIV-1的gag、pol和gp41区设计3套引物,建立检测HIV DNA的多重nPCR和检测HIVRNA的多重RT-PCR方法;分别以建立的PCR方法和NASBA法对119例HIV阳性患者进行检测,比较2者的检测敏感性;将患者分为不同的病毒载量组,比较PCR方法在各组患者中的检测敏感性差异;使用建立的PCR方法对10例可疑急性感染者进行检测;扩增HIV-1膜区C2-C3段,对nPCR检测阳性的43例DNA样本进行亚型鉴定。结果多重nPCR的检测敏感度为97.5%(116/119),多重RT-PCR的敏感度为78.2%(93/119),2者的特异度均为100%(50/50),多重nPCR和多重RT-PCR的阳性预测值分别为97.5%(116/119)和78.2%(93/119),阴性预测值均为100%(50/50),准确性分别为98.2%和84.6%。经比较,多重nPCR和多重RT-PCR的检测敏感度都高于NASBA法。在病毒载量<103copy/mL的患者,nPCR的检测敏感性高于RT-PCR,在病毒载量在(103～104)copy/mL的患者及病毒载量≥104copy/mL的患者,2者的敏感性比较相近,均接近100%;检测的10例可疑急性感染者中,有5例患者的nPCR和RT-PCR检测结果阳性,抗体在随访过程中阳转,证实为HIV急性感染者;检测的43例DNA样本分属于B’亚型(37例),AE亚型(5例)和BC亚型(1例)。结论本课题组建立的PCR检测方法可以对我国主要流行的B’、AE和BC亚型病毒株得到很好的扩增效果;nPCR的检测敏感性受血浆RNA水平的影响相对较小,RT-PCR的检测敏感性受血浆RNA水平的影响相对较大;PCR方法可以用于HIV急性感染者的早期诊断。     

宏基因组测序辅助HIV感染者发热待查的病原学诊断1例报告

探讨宏基因组测序（metagenomic next-generation sequencing,mNGS）技术在发热待查中的诊断应用价值。发热待查一直是临床诊治难点,本研究中1例人类免疫缺陷病毒（HIV）感染者反复发热2+月,经涂片、5次外周血培养和1次骨髓培养、血清学和病理检查等常规病原微生物检测未发现病因。分别取淋巴结进行广谱聚合酶链式反应（polymerase chain reaction,PCR)检测和取外周血进行mNGS检测;广谱PCR和mNGS检测结果均确定病原菌为马尔尼菲篮状菌。追加外周血培养（并延长培养时间）最终获得菌株,并行全基因组测序。mNGS与之比较,全基因组序列查见耐药基因FLU1,但mNGS未能检出;全基因组序列显示该菌株与NCBI数据库中的已知菌株均没有亲缘关系,而mNGS因病原体数据量不够而不能做亲缘关系分析。本研究报告了1例应用mNGS诊断马尔尼菲蓝状菌感染的病例,表明mNGS能在辅助HIV感染者发热待查的病原学诊断中发挥重要作用,但mNGS的耐药基因检测和病原体溯源等功能尚需后续发展。     

肾移植患者人巨细胞病毒的检测

采用ＰＣＲ和ＥＬＩＳＡ方法对１１例肾移植者的巨细胞病毒（ＨＣＭＶ）感染情况进行了研究。在术前及术后的４周内，对其外周血及尿中病毒和血清中ＩｇＡ及ＩｇＭ抗体进行了连续检测。结果显示：有６例在移植后出现巨细胞病毒感染，感染率为５４％（６／１１）。血、尿中ＨＣＭＶＤＮＡ或ＨＣＭＶ抗体（ＩｇＡ或ＩｇＭ）阳性。提示：用ＰＣＲ和ＥＬＩＳＡ方法对肾移植病人的ＨＣＭＶ感染情况进行监测，对预防和治疗ＨＣＭＶ感染和提高肾移植的成功率有着重要的意义。     

孕早期原发性巨细胞病毒感染对胎、婴幼儿的影响

采用酶联免疫吸附试验及多聚酶链反应技术对孕早期肯定为原发性巨细胞病毒感染的20例孕妇进行前瞻性研究，并与中、晚期诊断为再发感染及既往感染的孕妇胎、婴幼儿进行对照研究，发现原发感染者其宫内垂直传播率为75．0％（15／20），活产新生儿先天性感染率为66．67％（10／15），胎儿宫内发育迟缓发生率为20．0％，死胎、畸形的出现率分别为5．0％（1／20）、10．0％（2／20），均明显高于再发感染及既往感染组。采用贝利发育量表对10例原发感染组、13例再发感染组及34例既往感染组孕妇所出生的婴幼儿进行检测，结果表明原发感染组的婴幼儿智力发育指数明显低于再发感染及继往感染组（P＜0．05），精神运动发育指数三者之间差异无显著意义（P＞0．05）。提示孕早期原发性巨细胞病毒感染对胎、婴幼儿影响多而且严重，孕早期原发性巨细胞病毒感染的筛查极为重要。     

聚合酶链反应诊断新型隐球菌脑膜脑炎50例

目的 探讨聚合酶链反(PCR) 新型隐球菌脑膜脑炎快速诊断的临床标准。方法 应用聚合酶链反应检测50例临床脑脊液标本中的新型隐球菌，同时用直接涂片墨汁复杂、真菌培养进行对比检测。结果 在50例患者临床脑脊液标本中，PCR检测阳性11例；直接涂片墨汁复染阳性7例阳性；真菌培养阳性9例。结论 用PCR方法具有简便、快速、特异、敏感的特点，从脑脊液标本中直接检测新型隐球菌，提供病原学诊断依据，对新型隐球菌脑膜脑炎的快速诊断具有重要的临床意义。     

Use of interventions for reducing mother-to-child transmission of HIV in Australia

OBJECTIVE: To describe the extent and outcome of use of interventions for reducing the risk of HIV transmission from mother to child in Australia. DESIGN: National surveillance for perinatal exposure to HIV. PARTICIPANTS AND SETTING: Notified cases of HIV infection in women in Australia and their perinatally exposed children, 1982-1999. OUTCOME MEASURES: Trends over time in use of interventions (antiretroviral therapy in pregnancy, elective caesarean delivery and avoidance of breastfeeding) and perinatally acquired HIV infection. RESULTS: By 31 March 2000, 204 children were reported as having been born in 1982-1999 to 162 women whose HIV infection had been diagnosed by 31 December 1999. The child's HIV infection status was established for 182 (89.2%); the mother's HIV infection was diagnosed antenatally in 91 of these cases (50%). Among women diagnosed antenatally, use of elective caesarean delivery and antiretroviral therapy in pregnancy increased significantly, from 3% and 14% by women whose children were born in 1982-1993, to 21% (P=0.01) and 88% (P<0.001), respectively, by women whose children were born in 1994-1999. Most women (95%) diagnosed antenatally avoided breastfeeding their children. The percentage of infected children born to women diagnosed antenatally declined from 26% among children born in 1982-1993 to 19% among those born in 1994-1999. The percentage of infected children was significantly lower among those whose mothers used antiretroviral therapy in pregnancy (11% versus 36%; P=0.03). CONCLUSION: Antiretroviral use in pregnancy, elective caesarean delivery and avoidance of breastfeeding have been effective interventions for reducing the risk of mother-to-child HIV transmission in Australia. While the rate of perinatal HIV transmission has declined, it remains high in comparison with rates reported from other industrialised countries.     

Comparison of Tzanck smear, viral culture, and DNA diagnostic methods in detection of herpes simplex and varicella-zoster infection.

OBJECTIVE--To compare Tzanck smears, viral cultures, and DNA diagnostic methods using the polymerase chain reaction (PCR) in detection of herpes simplex virus (HSV) or varicella-zoster virus (VZV) infection in clinically suspected cases. DESIGN--A 12-month trial comparing PCR with viral cultures and Tzanck smears in patients with clinically suspected HSV or VZV infection. SETTING--Both ambulatory and hospitalized patients were recruited from a tertiary referral center and the Miami (Fla) Veterans Affairs Medical Center. PATIENTS--Convenience samples of patients clinically suspected to have HSV (n = 48) or VZV (n = 35). To be included in the final analysis patients needed to have a positive Tzanck smear, viral culture, or PCR result. Patients who were clinically suspected to have HSV but had VZV by viral culture or PCR were analyzed in the VZV group. Similarly, patients who were clinically suspected to have VZV, but had HSV by viral culture or PCR were analyzed in the HSV group. Seventy-seven patients were available for final analysis: HSV (n = 30), VZV (n = 32), and 15 control cases who did not have evidence of viral infection. RESULTS--For HSV, PCR detected HSV DNA sequences in 73% of stained smears and 83% of unstained smears. For VZV infection, VZV DNA sequences were detected in 88% of stained smears and 97% of unstained smears. Viral DNA sequences were not detected in the 15 control cases. Viral cultures were positive in 83% and 44% of HSV and VZV cases, respectively. The Tzanck smear was positive in 60% and 75% of HSV and VZV cases, respectively. CONCLUSIONS--PCR is a reliable method for detecting HSV and VZV DNA sequences from single stained and unstained Tzanck smears. It is clearly superior to viral culture in identifying VZV infection and is equivalent to conventional culture techniques in identifying cases of HSV.     

The role of DNA amplification technology in the diagnosis of infectious diseases

Nucleic acid amplification and detection methods developed in the past decade are useful for the diagnosis and management of a variety of infectious diseases. The most widely used of these methods is the polymerase chain reaction (PCR). PCR assays can detect rapidly and accurately the presence of fastidious and slow-growing microorganisms, such as Chlamydia, mycoplasmas, mycobacteria, herpesviruses and enteroviruses, directly from clinical specimens. Commercial PCR assays for the diagnosis of tuberculosis and genital C. trachomatis infection are now routinely used in many diagnostic laboratories. Assays have also been developed that can detect antimicrobial resistance and are used to identify the cause of infection by organisms that cannot be cultivated. The value of viral load measurement by nucleic acid amplification in the management of patients with HIV infection or hepatitis C has also been well established. However, evaluations of this technology for rapid microbial diagnosis have generally been limited by small samples, and the cost of these assays may be as high as Can$125 per test. As nucleic acid amplification methods continue to evolve, their role in the diagnosis and management of patients with infectious diseases and their impact on clinical outcomes will become better defined.     

聚合酶链反应技术诊断中枢神经系统肠道病毒感染

目的：探讨肠道病毒（ＥＶ）在中枢神经系统感染中的致病情况,建立一种检测ＥＶ感染的方法。方法：采用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）和病毒分离技术检测４０种ＥＶ标准毒株和４６例无菌性脑膜炎、脑炎病人脑脊液（ＣＳＦ）标本。     

HIV母婴阻断及其婴儿预防接种不良反应监测报告

摘要：目的评估实施HIV母婴阻断措施婴儿群对预防接种的耐受性。方法为HIV感染孕妇提供综合性母婴阻断措施,将其婴儿设为观察组,母亲mV阴性婴儿为对照组,对2组婴儿的预防接种不良反应（AEFI）进行统计分析。结果H1V感染孕妇经孕期规范抗病毒治疗后母婴阻断效果明显,其婴儿感染率远低于未治疗者。观察组98名、对照组100名婴儿共接种17种疫苗2428人次。观察组和对照组发生AEFI分别为2．9％和2．1％（P〉0．05）,其中一般反应分别占75．6％、78.11％,异常反应分别占4．4％和0,偶合症分别占20．0％、21．9％。全身反应以发热为主,局部反应以接种部位无菌性脓肿为主,2组婴儿均无严重预防接种不良反应事件,也没有因接种卡介苗引起的播散性卡介苗病例发生。结论HIV阳性孕妇经综合性HIV母婴阻断措施后其所娩婴儿按国家计划免疫程序进行预防接种是安全的．     

生殖器疱疹实验诊断方法的临床应用探讨

目的 探讨生殖器疱疹实验诊断方法的临床应用价值.方法 采用细胞培养法对82例临床诊断为典型生殖器疱疹的患者进行人类单纯疱疹病毒(HSV)分离培养,并同时进行聚合酶链反应(PCR)检查.结果 两种方法的HSV检出率分别为58.5%和92.6%;培养法作为金标准特异性高,对早期典型水疱检出率高达89.7%;PCR与培养法相比敏感性高,尤其对糜烂或溃疡标本阳性检出率明显高于培养法.结论 培养法适于早期检测,用于生殖器疱疹诊断特异性高;PCR操作具有简便、快速及敏感性高等特点,可用于不同皮损的检测.     

定量PCR检测对侵袭性真菌感染肺组织标本病原真菌的诊断价值

目的探讨应用定量聚合酶链反应(polymerase chain reaction,PCR)检测技术诊断侵袭性真菌感染(invasive fungal infection,IFI)肺组织标本病原真菌的临床意义。方法收集2010年6月至2016年6月间我院行手术切除且病理诊断为真菌感染的肺组织标本33例和同期非真菌感染的正常肺组织标本10例为研究对象,设计针对真菌的通用引物和针对隐球菌属和曲霉菌的特异性引物,采用PCR技术和双脱氧链终止法(Sanger法测序)检测石蜡包埋肺切除组织中的真菌。结果实验组33例真菌感染的肺组织标本PCR检测结果均为阳性,与病理诊断相符,Sanger测序可以鉴定真菌到种别。PCR检测与Sanger测序敏感性差异无统计学意义(P>0.05),PCR检测与Sanger测序特异性差异无统计学意义(P>0.05);PCR检测与镜检的种属鉴定率差异有统计学意义(P<0.05),PCR检测与Sanger测序的种属鉴定率差异无统计学意义(P>0.05)。结论定量PCR检测用于诊断侵袭性真菌感染肺组织标本敏感性高、特异性好,具有一定参考意义。     

The impact of a HIV prevention of mother to child transmission program in a nigerian early infant diagnosis centre

Background:

Mothers infected with human immunodeficiency virus (HIV) can transmit the virus to their babies in utero, intrapartum or postpartum through breastfeeding. Maternal to child transmission can be prevented through administration of antiretroviral drugs to mother and child, and through restriction of breastfeeding. This study evaluated the effectiveness of prevention of mother-to-child transmission (PMTCT) activities in reducing the incidence of HIV infection among exposed babies at the National Hospital Abuja, Nigeria.

Materials and Methods:

Early infant diagnosis laboratory records of 515 exposed babies aged below 18 months who had polymerase chain reaction (PCR) test between January 1^st 2011 and December 31^st 2012 were reviewed. The details of antiretroviral (ARV) therapy commencement for mother and baby, infant feeding choices, mode of delivery and HIV test results were analysed.

Results:

Of the 515 samples tested, 36 (7.0%) were found to be positive. The mean age of exposed children tested was 4 months. Highest prevalence was among children in the age group 6-18 months (16.1%). There was statistically significant association between HIV positive results and age. (P = 0.0000). If the mother and child pairs received ARVs, the prevalence was 1.3%, whereas if the mother only received ARV, then the prevalence was 4.6%, and when only the child received ARV the prevalence was 20.0%. When neither the mother nor the child received ARVs, the prevalence was 66.7%.

Conclusion:

There was a high prevalence of HIV among exposed children in our setting, especially if the mother and child pairs did not receive any form of antiretroviral prophylaxis. This further emphasises the usefulness of ARVs as the single most important intervention in PMTCT. Therefore, there is need to expand antiretroviral coverage, ensure access of the PMTCT program, and provide effective services to support infected children.     

肺炎患儿肺炎支原体感染的实验室检测

目的：探讨肺炎患儿肺炎支原体（ＭＰ）感染的实验室检测方法。方法：使用聚合酶链反应（ＰＣＲ）技术检测６７６例肺炎患儿ＭＰ的感染,其中１３９例同时采用支原体培养进行检测;１１２例做间接血凝试验（ＩＨＡ）。结果：ＰＣＲ法对ＭＰ感染的检出率明显高于培养法（Ｐ〈０．０１）,双份血清ＩＨＡ的检测率和ＰＣＲ法比较无显著性差异（Ｐ〉０．０５）,单份血清ＩＨＡ的检出率明显低于ＰＣＲ法（Ｐ〈０．０１）。结论：ＰＣＲ法     

逆转录聚合酶链反应方法检测新生儿病房柯萨奇B_3病毒感染

目的:探讨逆转录聚合酶链反应(RT-PCR)方法在检测柯萨奇病毒B3(CoxB3)感染中的作用。方法:采用RT-PCR方法对北京某医院31对(62例)疑似有母婴传播性CoxB3感染的新生儿和母亲血样进行CoxB3病毒的RNA检测。结果:62例样本经血清学检测,只有1例新生儿CoxB3病毒抗体阳性,阳性率为1.61%。经RT-PCR方法检测,62例血样中有12例检测结果为阳性,阳性率为19.35%;其中4对母、婴均阳性,2对新生儿阳性、母亲阴性,还有2对母亲阳性、新生儿阴性。其余23对母婴均为阴性。结论:RT-PCR法对于早期CoxB3感染检测的阳性率优于血清学检测方法,对确定病毒的传播途径有参考价值,同时可以早期预防病毒感染在新生儿中的爆发。RT-PCR法具有快速、敏感、特异性强等特点,是快速、有效检测CoxB3病毒感染的诊断方法之一。     

PCR法快速诊断急性重症胰腺炎早期合并细菌感染

目的探讨以16srRNA基因为基础的细菌聚核酶链反应（PCR）法诊断急性重症胰腺炎早期合并细菌感染的价值。方法收集17例急性重症胰腺炎患者（女13例，男4例，年龄17～79岁）不同时期、不同部位标本45份，其中17份来自细针穿刺（FNA），28份为术中标本。获取胰周渗液后，行细嚣通用引物PCR法检测。并与常规细菌培养结果进行比较。结果10例最终诊断为胰腺感染，并行手术治疗；7例最终诊断为胰腺未感染，5例经保守治疗治愈，2例死亡。常规培养阳性检出率为84．4％；细菌PCR法阳性检出率为80％。PCR、培养诊断胰腺感染的敏感性分别为90％（9／10）、100％（10／10），特异性均为100％（7／7）。两者耗时分别为5h、3d。结论B超引导细针穿刺安全可行；PCR法能快速、准确地诊断急性重症胰腺炎早期合并细菌感染，为手术治疗提供可靠依据。     

HIV重叠肝炎病毒感染者血清IL-18和IL-10水平分析

目的：探讨细胞因子在HIV—HBV／HCV重叠感染中的意义。方法：采用ELIsA法检测血清IL—18和IL—10的水平，荧光定量PCR计数测定HIV—RNA滴度，流式细胞仪检测CD4^ T细胞。结果：HIV感染后，血清IL—18显著高于正常对照(P＜0．05)。随着疾病的进展，IL—18逐渐上升而IL—10逐渐下降。重叠感染组血清IL—18和IL—10显著高于单纯HIV感染组。结论：肝炎病毒感染是影响HIV感染者血清细胞因子水平的一个协变量，上调IL—18和下调IL—10可能起到改善重叠感染的预后的作用。