输血后乙型和丙型肝炎病毒感染的前瞻性调查

目的 了解输血所致ＨＢＶ和ＨＣＶ感染现状及ＨＢｓＡｇ抗ＨＣＶ和ＡＬＴ筛检供血的效果。方法 对１３８例输血者输血前，后１，３，６，９个月血清标本及其供血检测ＨＢｓＡｇ，抗ＨＢｓ，抗ＨＢｃ，抗ＨＣＶ和ＡＬＴ，并对部分血清标本用ＰＣＲ及巢式ＲＴ－ＰＣＲ法检测ＨＢＶＤＮＡ和ＨＣＶＲＮＡ，结果和结论 输血后ＨＢＶ和ＨＣＶ感染率分别为１．４％（２／１３８）和３４．８％（４８／１３８），输血后乙型和丙型肝炎发生     

佛山地区存在经输血传播病毒(TTV)感染

我们应用Ｎｅｓｔｅｄ－ＰＣＲ法检测了４５例甲到戊型标志物均阴性的肝炎患者和４４例献血员血清中的ＴＴＶＤＮＡ。资料与方法：４５例肝炎患者均为我院传染科或门诊病人,血清经ＥＬＩＳＡ法检测抗－ＨＡＶ,ＨＢｓＡｇ,ＨＢｃＡｂ,抗－ＨＣＶ,抗－ＨＥＶ均阴性,ＰＣＲ检测ＨＢＶＤＮＡ,ＨＣＶＲＮＡ均阴性。４４例某院献血员检测抗－ＨＡＶ,ＨＢｓＡｇ,ＨＢｃＡｇ,ＨＢｃＡｂ,抗－ＨＣＶ,抗－ＨＥＶ为阴性,肝功能正常。抽提标本血清ＤＮＡ进行Ｎｅｓｔｅｄ－ＰＣＲ：以ＲＤ０３７和ＲＤ０３８为引物进行第一次ＰＣＲ,再以Ｒ…     

PCR结合地高辛标记探针杂交检测乙型肝炎病毒C基因启动子

应用聚合酶链反应 (ＰＣＲ)结合地高辛标记探针杂交方法检测乙型肝炎病毒Ｃ基因启动子 (ＨＢＶ .ＣＰ)。对ＰＣＲ法扩增的ＨＢＶＤＮＡ直接测序进行ＤＮＡ同源性分析 ,并运用探针杂交对结果作进一步鉴定。应用ＰＣＲ法扩增 2 0份患者血清 ,阳性率达 95% ( 19/ 2 0 ) ,同时运用探针杂交作进一步检测 ,与ＰＣＲ结果符合率达 90 %以上 ,并选择中度、重度乙型肝炎患者血清和 ｐＧＥＭ .7Ｚ ＨＢＶ质粒扩增的ＨＢＶ .ＣＰ各 1份分别进行测序 ,与报告基因的同源性分别为 72 %、66.5%和 90 %。建立了特异敏感的检测ＨＢＶ .ＣＰ的ＰＣＲ方法 ,可用…     

巢式PCR检测血液病血清中HCV RNA

巢式ＰＣＲ检测血液病血清中ＨＣＶ　ＲＮＡ仉红刚，张秋菊，王景明通过输血途径感染丙型肝炎病毒（ＨＣＶ）是血液病治疗中的一个长期潜在的并发症。为难确、及时地检测ＨＣＶ的近期感染，采用巢式ＰＣＲ技术对４６例血液病患者血清中ＨＣＶＲＮＡ进行了检测研究，现将有...     

多重和套式聚合酶链反应检测血液,腹膜透析患者血清中HBV D…

利用免疫学和聚合反应（ＰＣＲ）方法，对３５例血液透析（血透），１４例腹膜透析（腹透）患者的８４份血清进行了检测。血透组抗－ＨＣＶ阳性率８２．９％，ＨＣＶ ＲＮＡ ＰＣＲ阳性率５１．４％；腹透组抗－ＨＣＶ阳性率仅７．１％，ＨＣＶ ＲＮＡ ＰＣＲ均阴性。ＨＢｓＡｇ和（或）ＨＢｅＡｇ在二组的阳性率分别为２５．７％和２１．４％；ＨＢＶＤＮＡＰＣＲ阳性率分别为４５．７％和６４．２％。透析患者ＨＣＶ和ＨＢＶ感     

套式及免疫套式聚合酶链反应检测乙型肝炎表面抗原阴性肝病患者血清中乙型肝炎病毒DNA

为了更准确、简便而又快速地了解乙型肝炎表面抗原（ＨＢｓＡｇ）阴性肝病患者的病因，建立了套式和免疫套式聚合酶链反应（ＰＣＲ）检测血清中乙型肝炎病毒（ＨＢＶ）ＤＮＡ的技术，结合丙型肝炎病毒（ＨＣＶ）感染指标的检测，对ＨＢｓＡｇ阴性肝病患者的病因进行了研究。发现套式ＰＣＲ能将单次ＰＣＲ的敏感性稳定地提高１０００倍；免疫套式ＰＣＲ可检测到０．１～０．０１ｐｇ／Ｌ水平。检测ＨＢｓＡｇ阴性肝硬化２２例（Ａ组）、ＨＢｓＡｇ阴性慢性肝炎１３例（Ｂ组）、ＨＢｓＡｇ阴性和乙型肝炎病毒核心抗体（抗－ＨＢｃ）阳性正常对照组３０例（Ｃ组）及ＨＢｓＡｇ（＋）／ＨＢｅＡｇ（－）肝硬化患者１２例（Ｄ组），分别有４５．５％、３０．８％、１３．３％和１００％患者血清中ＨＢＶＤＮＡ阳性。ＨＢＶＤＮＡ在一些抗－ＨＢｓ（＋）肝病患者和所谓健康人的血清中也存在。Ａ、Ｂ两组检出有ＨＢＶ和（或）ＨＣＶ感染患者分别占８１．８％和５３．８％。提示套式和免疫套式ＰＣＲ是简便、快速而又高度敏感的检测方法；ＨＢＶ感染可能是引起ＨＢｓＡｇ阴性慢性肝病的重要原因，且ＨＢｓＡｇ阴性肝病病因大多与病毒感染有关；应该重新认识自然感染者血清中抗－ＨＢｓ的临床意义。     

多重和套式聚合酶链反应检测血液、腹膜透析患者血清中HBV DNA和HCV RNA

利用免疫学和聚合酶链反应（ＰＣＲ）方法，对３５例血液透析（血透）、１４例腹膜透析（腹透）患者的８４份血清进行了检测。血透组抗－ＨＣＶ阳性率８２．９％，ＨＣＶＲＮＡＰＣＲ阳性率５１．４％；腹透组抗－ＨＣＶ阳性率仅７．１％，ＨＣＶＲＮＡＰＣＲ均阴性。ＨＢｓＡｇ和（或）ＨＢｅＡｇ在二组的阳性率分别为２５．７％和２１．４％；ＨＢＶＤＮＡＰＣＲ阳性率分别为４５．７％和６４．２％。透析患者ＨＣＶ和ＨＢＶ感染率显著高于一般人群。血、腹透两组ＨＣＶ感染率相差非常显著，血透患者抗－ＨＣＶ和ＨＣＶＲＮＡＰＣＲ阳性高于腹透患者，表明血透过程在丙型肝炎传播方面起重要作用。作者还采用多重和套式ＰＣＲ方法，将ＨＢＶＤＮＡ和ＨＣＶＲＮＡ在合并的逆转录和ＰＣＲ扩增系统中，连续进行逆转录和第一轮多重ＰＣＲ扩增，再进行第二轮多重ＰＣＲ，具有特异、敏感、简便、快速和经济等特点，适于临床应用。     

多重聚合酶链反应检测乙型肝炎和丙型肝炎病毒

在同一反应体系中同时快速检测乙型肝炎病毒（ＨＢＶ）和丙型肝炎病毒（ＨＣＶ），探讨多重聚合酶链反应（ＰＣＲ）在临床中的应用价值。方法自行设计了ＨＢＶ和ＨＣＶ各一对引物，应用多重ＰＣＲ对临床标本进行检测。结果５例ＨＢｓＡｇ和ＨＣＶ抗体均阳性者，其ＨＢＶＤＮＡ和ＨＣＶＲＮＡ均阳性；２０例单ＨＢｓＡｇ阳性标本，１１例单ＨＢＶＤＮＡ阳性，５例ＨＢＶＤＮＡ和ＨＣＶＲＮＡ均阳性；２０份单ＨＣＶ抗体阳性者，ＨＣＶＲＮＡ均阳性，３例合并ＨＢＶＤＮＡ阳性；１０例慢性非甲非乙型肝炎（ＮＡＮＢ）抗ＨＣＶ阴性肝炎标本８份ＨＣＶＲＮＡ阳性，１例ＨＢＶＤＮＡ和ＨＣＶＲＮＡ阳性。结论多重ＰＣＲ一次反应即可完成多次常规ＰＣＲ检测，是一种值得临床应用的方法。     

乙型用丙型肝炎病毒感染与肝细胞癌

为探讨乙型和丙型肝炎病毒（ＨＧＢＶ和ＨＣＶ）感染与肝细胞癌（肝癌）发生的关系采用ＥＬＩＳＡ和聚合酶链反应（ＰＣＲ）对沈阳地区１１７例肝癌、１０７便肝硬化和４５ 析患者血清进行了ＨＢＶ和ＨＣＶ血清标志及ＨＢＶＤＮＡ和ＨＣＶ ＲＮＡ检测，并采用性片段长度多态性对其中７３便ＨＣＶ ＲＮＡ阳性血清进行了ＨＣＶ基因分型。结果，肝癌组ＨＢＶ感染率（６０．７％）显著高于ＨＣＶ感染率（３３．５％，Ｐ〈０．０１），     

28例HCV RNA阳性血清聚合酶链反应产物的酶切分析

为了探讨广州地区ＨＣＶ感染的基因型，作者对广州地区不同人群中１０５例抗－ＨＣＶ阳性血清用逆转录聚合酶链反应（ＴＲ－ＰＣＲ）进行了ＨＣＶ ＲＮＡ的检测，对其中８２例ＨＣＶ ＲＮＡ阳性患者血清做了ＰＣＲ产物的限制性内切片段长度多态性（ＲＦＬＰ）分析，结果显示：ＨＣＶⅡ型感染７８例（９２．１％），ＨＣＶⅢ型１例，ＨＣＶⅡ／Ⅲ型混合感染３例，４例ＨＣＶⅢ型感染均是肝病患者，初步表明广州地区ＨＣＶ感染大多数     

Hepatitis delta virus RNA in serum of patients with chronic delta hepatitis

We studied 28 patients with chronic delta hepatitis for the presence of hepatitis delta virus (HDV) RNA in serum. The hot start polymerase chain reaction (PCR) method, in which the reaction begins at 60–80° C, showed a higher sensitivity than conventional PCR reaction. Additionally, the presence of hepatitis B (HBV) and C virus (HCV) infections were determined by PCR. HDV RNA was detected in 26 patients (93%), HBV DNA in 22 (79%), and HCV RNA in only one. Detection of HDV RNA correlated very well with detection of hepatitis delta antigen by immunostaining in the liver. In six patients HDV RNA was detectable despite the absence of HBV DNA in serum, suggesting that high levels of HBV are not required for HDV replication. Of 29 control patients with chronic hepatitis B without antibody to HDV, none had detectable HDV RNA, while all had HBV DNA in serum. Detection of HDV RNA with PCR proved highly sensitive and specific, demonstrating that virtually all patients with chronic HDV infection had ongoing viral replication.     

合并乙型肝炎病毒感染对丙型肝炎病毒核心抗原检测的影响

目的 探讨合并HBV感染对慢性HCV感染者血清丙型肝炎病毒核心抗原(HCVcAg)检出情况的影响. 方法 收集2005年12月-2009年10月慢性丙型肝炎患者和HBV/HCV合并感染者资料,检测血清HCVcAg和HCV RNA,对后者血清进行HBV DNA、HBeAg检测,分析HCVcAg检出率与HBeAg、HBV DNA定量检测的关系.用独立两组多分类的X2检验方法进行统计学分析. 结果 共收集88例慢性丙型肝炎患者和62例HBV/HCV合并感染者资料,血清HCVcAg的检出率分别为72.7％(64/88)和38.7％ (24/62),两者比较,x2= 17.358,P＜0.01,差异有统计学意义.HCV RNA检出率分别为81.8％ (72/88)和53.2％ (33/62),两者比较,x2=20.110,P＜0.01,差异有统计学意义.62例HBV/HCV合并感染者血清中,HBeAg阳性和HBeAg阴性感染者HCVcAg检出率分别为28.6％ (12/42)和60.0％ (12/20),两者比较,x2=5.641,P=0.011,差异有统计学意义.HCV RNA阳性率分别为42.9％ (18/42)和80.0％ (16/20),两者比较,X2=7.547,P＜ 0.01,差异有统计学意义.HBV DNA阳性和阴性时HCVcAg检出率分别为39.1％ (18/46)和37.5％ (6/16),两者比较,P＞0.05,差异无统计学意义.与单纯HCV感染者血清HCVcAg检出率72.7％ (64/88)比较,HBeAg阴性合并感染者为60.0％ (12/20),x2=1.266,P=0.261,差异无统计学意义;HBV DNA阴性合并感染者为37.5％ (6/16),x2=7.635,P＜0.01,差异有统计学意义.结论 HBV/HCV合并感染时HCVcAg检出率较低,可能是由于HBeAg抑制HCV的复制,从而减少HCVcAg的表达所致.     

肝癌组织中丙型和乙型肝炎病毒基因状况研究

采用逆转录巢式ＰＣＲ技术检测了５１例ＨＣＣ患者的肝癌及癌周肝组织中ＨＣＶＲＮＡ正负链，同时以巢式ＰＣＲ技术检测了这些组织中的ＨＢＶＤＮＡ。结果，１１．８％（６／５１）检出组织中ＨＣＶＲＮＡ正链：其中５例在癌周肝组织，２例在癌组织中检测出ＨＣＶＲＮＡ负链，由于负链ＲＮＡ为ＨＣＶ的复制中间体，不释放到细胞外，其检出ＨＣＶ感染与ＨＣＣ关系的研究提供了进一步的病因证据，组织中ＨＢＶＤＮＡ检出率高达９２．２     

血液透析患者乙、丙型肝炎感染的状况

目的研究血液透析患者乙、丙型肝炎感染状况及感染途径。方法对４９例维持性血液透析患者用套式逆转录聚合酶链反应方法（ＲＴＰＣＲ）检测了乙型肝炎病毒脱氧核糖核酸（ＨＢＶＤＮＡ）及丙型肝炎病毒核糖核酸（ＨＣＶＲＮＡ），用第二代酶联免疫吸附法（ＥＬＩＳＡ）检测丙型肝炎抗体、乙型肝炎标志物（ＨＢＶＭ）。结果透析患者乙型肝炎感染率５３．１％，丙型肝炎感染率６９．４％。输血组乙、丙型肝炎感染率大于未输血组；透析程２４个月以上者丙型肝炎感染率大于透析１２个月以内者（Ｐ＜００５）。ｌｏｇｉｓｔｉｃ多元回归分析显示，透析年限的增加是丙型肝炎感染的主要因素，提示透析程的危险度大于输血危险度。２０份血站提供的血制品有３份ＨＣＶＲＮＡ阳性，２０份复用透析器经消毒处理后检测出２份ＨＣＶＲＮＡ阳性，９例工作人员ＨＣＶＲＮＡ及ＨＢＶＤＮＡ均阴性。结论加强透析室的管理及工作人员的防护，减少输血及透析器的复用，对减少透析中乙、丙型肝炎感染至关重要     

维持血液透析的尿毒症病人乙型丙型肝炎病毒感染情况研究

目的　了解北京地区 (东城、宣武、朝阳区 )接受规律性血液透析患者 ,乙型肝炎病毒 (HBV)和丙型肝炎病毒 (HCV)的感染情况及其相关因素。方法　 1998年 3～ 12月在北京协和医院、朝阳医院等 4家医院血液净化中心 ,长期维持血液透析的尿毒症患者 2 2 5例 ,血透中心工作人员及健康献血者 5 0例为对照组。分别用PCR法和高敏PCR法检测HBVDNA、HCVRNA ,ELISA法检测乙肝两对半及丙肝抗体 ,并分析其与透析时间、输血、肝功能损害的关系。结果　 2 2 5例血液透析病人中 ,HCVRNA阳性 37例 (16 4 %) ;HBVDNA阳性 3例(1 33%)。多元回归分析表明 :输血和透析时间是丙型肝炎感染的危险因素。共有 3 0 %(3/ 99)的病人同时感染乙肝和丙肝 ,均有肝功能的损害和临床症状 ,8 1%(8/ 99)HBcAb阳性患者同时合并HCV感染。结论　血液透析病人乙肝和丙肝感染远高于对照组 ,透析时间和输血次数是丙肝感染的危险因素 ,HBV和HCV同时感染问题值得重视。     

Contribution of HEV and HCV in causing fulminant non-A, non-B hepatitis in western India

Summary. During 1990, 38 patients with fulminant non-A, non-B hepatitis (NANB) died in Government Medical College Hospital, Aurangabad. Serum samples from these patients were tested for antibodies to hepatitis C virus (anti-HCV) and IgM antibodies to hepatitis E virus (IgM-anti-HEV). All samples were also subjected to polymerase chain reaction (PCR) for the detection of HBV DNA, HCV RNA and HEV RNA. None of the patients had circulating anti-HCV antibodies; three had HCV RNA. Based on anti-HEV-IgM positivity 14 patients (37%) could be diagnosed as suffering from hepatitis E. None was positive for HEV RNA. In the absence of serological markers, HBV DNA was present in three cases. None of the HBV DNA positive patients had anti-δ antibodies. Dual infections (HBV with HEV, and HBV with HCV) were seen in two cases. The aetiology of half of the NANB cases could not be assigned to the known hepatitis viruses using current techniques.     

Prevalence of occult hepatitis B & C in HIV patients infected through sexual transmission.

BACKGROUND: Prevalence of Hepatitis B virus (HBV) and Hepatitis C virus (HCV) markers including active and occult infection has not been described in diverse cohorts among HIV-infected patients in India. Earlier studies have explained the role of HBV/HCV co-infection in cohorts of injection drug users (IDUs) but the sexual co-transmission of HBV/ HCV is not completely understood. OBJECTIVE: The objective of this study was to assess the prevalence of occult HBV & HCV infection in HIV positive sexually acquired transmission risk group. MATERIALS AND METHODS: 58 sexually acquired HIV positive patients were taken up for the study of occult HBV/HCV co-infection. Data on demographics, sexual behaviour, sexually transmitted diseases (STD), medical history, laboratory tests viz., serum ALT and CD4 count were recorded. HBV serology included HBsAg, anti HBs, IgG anti HBc and HBV DNA (PCR). HCV serology included anti HCV & HCV RNA (RT-PCR). RESULTS: Occult HBV infection (HBV DNA) was observed in 12.2% (7/58 with HBsAg -ve and IgG anti HBc +ve subjects) while an overall prevalence of HBV DNA was 13.7% (12% occult & 1.7% in HBsAg+ve patients). Out of 58 HIV positive patients 29.3% demonstrated reactivity for any marker of past or current HBV infection. (HBsAg 1.7%, anti HBs 10.3% anti HBc IgG 17.2%). 4/58 (6.8%) revealed anti HCV positivity along with HCV RNA positivity by RT-PCR while 6/58 (10.3%) individuals revealed an occult HCV infection (anti HCV negative). The overall HCV RNA prevalence was 17.2%. 2 out of 58 (3.4%) individuals were positive for occult infection of both HBV DNA & HCV RNA (Triple infection HIV/HBV/ HCV). The HBV/HCV co-infected group (n = 18) showed a significantly high ALT (114.3 + 12.3 U/I) & low CD4 count (202.5 + 33.7 cells/mm3). The percent prevalence of HBV/ HCV co-infection was higher in the illiterate group, in men less than 30 years of age, and in those who were married and exhibited polygamous activity. CONCLUSIONS: The study demonstrated that in HIV infected patients testing only serological viral markers like HBsAg, antiHBcIgG & anti HCV, fails to identify the true prevalence of co-infection with HBV & HCV. Qualitative PCR for HBV DNA & HCV RNA detects co-infection in patients who are negative for serological markers. Also, in subjects who had only a sexual risk factor for parenterally transmitted infections, HIV may enhance the sexual transmission of HBV and HCV.     

Profound suppression of chronic hepatitis C following superinfection with hepatitis B virus.

BACKGROUND: Hepatitis B virus (HBV) and hepatitis C virus (HCV) share similar transmission routes; thus, coinfection is frequent. The consequences of acute HBV infection in patients with chronic hepatitis C are unknown. METHODS: We describe a 47-year-old male with chronic hepatitis C who acquired HBV and then spontaneously and apparently completely cleared HCV but developed chronic hepatitis B. Five serum samples collected over 14 months and lymphoid cells obtained after acquiring HBV were tested for HCV and HBV by both standard assays and ultra-sensitive polymerase chain reaction/nucleic acid hybridization (PCR/NAH) (sensitivity approximately 2 IU/ml). RESULTS: After superinfection with HBV, HBV surface antigen-positive chronic hepatitis developed with readily detectable HBV DNA. All sera collected after acquisition of HBV, which tested negative for HCV RNA by standard laboratory assay, were positive for HCV genomes when analysed by PCR/NAH. Peripheral lymphoid cells carried HBV DNA and covalently closed circular DNA, but were negative for HCV. CONCLUSIONS: This is the first reported case of profound suppression of chronic hepatitis C after superinfection with HBV and establishment of chronic hepatitis B. It is hypothesized that HBV infection precipitated generalized and/or virus-specific cellular immune responses that profoundly suppressed HCV replication and yet failed to inhibit progression to chronic hepatitis B.     

慢性乙型肝炎患者外周血单个核细胞内HBV RNA、HBV DNA的检测及其与血清HBV DNA的关系

目的:探讨慢性乙型肝炎(慢乙肝)患者外周血单个核细胞(PBMC)中HBV感染情况及复制状态,以及与血清HBV DNA的关系.方法:应用PCR技术特异性,选择地检测慢乙肝患者PBMC中HBV RNA、HBV DNA,并与血清HBV DNA进行比较.结果:在慢乙肝患者PBMC中能检测出HBV RNA及HBV DNA,阳性率分别为38.78%及77.55%,且两者的检出率有一致性.当PBMC中HBV RNA阳性时,HBV DNA均为阳性(100%),而当HBV DNA阳性时,部分病例可表现为HBV RNA阴性,两者阳性符合率为50%.血清中HBV DNA阳性率为71.42%.PBMC中HBV RNA的阳性率与血清中HBV DNA的阳性符合率为63.33%,血清中HBV DNA与PBMC中HBV DNA阳性符合率为76.32%.结果提示:HBV能感染PBMC,在部分患者存在着活动性复制,这种复制表现为不同于肝细胞内的低水平复制.此外,HBV感染PBMC后也存在着非复制状态,表现为PBMC中HBV DNA阳性,但HBV RNA阴性.另外,当血中HBV DNA阴性时,少数病例PBMC仍可表现为HBV RNA及HBV DNA阳性.结论:HBV能够感染PBMC并存在活动性复制,可能是造成患者免疫功能紊乱、肝脏损害慢性化的重要原因之一.     

重叠乙型和丙型肝炎病毒感染的临床与病理分析

目的 观察HBV和HCV重叠感染的临床与病理,探讨HBV和HCV相互作用的特点.方法 收集226例慢性肝病患者的血清学指标,实时荧光定量PCR法测定HBV DNA和HCVRNA,ELISA检测HBV血清标志物、抗-HCV抗体.行肝穿刺活组织病理检查、免疫组织化学HBsAg、HBcAg和原位杂交HBV DNA、HCV RNA检测.计数资料比较采用X2检验或Fisher确切率检验.结果 HBV和HCV重叠感染的重度慢性肝炎患者比例为62.50%,高于HBV或HCV单独感染者的27.05%和30.56%(X2=14.70,P＜0.01).HBV感染组的血ALT、AST、TBil、DBil和Alb高于HBV和HCV重叠感染组和HCV感染组,差异均有统计学意义(X2=8.52,P＜0.05).重叠感染组和HBV感染组的血HBsAg与肝内HBsAg一致率比较,差异均有统计学意义(X2=15.60,P＜0.01).HBV和HCV重叠感染组血清HBV DNA阳性率为12.5%,低于HBV单独感染组的87.7%(X2=17.66,P＜0.01);而HBV和HCV重叠感染组HCV RNA阳性率为75.0%,低于HCV单独感染组的80.6%,差异无统计学意义(P＞0.05).结论 HBV和HCV重叠感染导致的肝损伤更明显.