献血者中TT病毒检测及其致病性探讨

目的　探讨TT病毒 (TTV)在输血过程中的传播及其致病性。方法　常规酚 氯仿法抽提血清病毒DNA ,设计位于TTVORF1保守区的两对引物 ,套式PCR扩增病毒DNA。ELISA检测血清HBsAg、抗 HAV、抗 HCV和抗 HIV。结果　健康献血者血清中TTV DNA检出率为 4 3 1 % (96 / 2 2 3)。受血者输血TTV DNA阳性血后 2周 ,血清TTV DNA转阳率为 6 3 6 % (1 4 / 2 2 )。 8周血清TTC DNA阳性率仍为 4 6 4 % (1 0 / 2 2 ) ,其中两对献血者和受血者之间血清TTV DNA同源性达 1 0 0 %。上述 2 2名受血者输血 2周后血清ALT、TB和DB正常 ,血清HBsAg、抗 HAV、抗 HCV和抗 HIV均阴性。随访其中 5例至 8周 ,无肝炎症状及体征 ,血清肝功能正常。结论　TTV可通过血液传播 ,但无明显致病性     

乙型肝炎病毒标志物及其DNA相关性及联合检测临床价值

目的探讨乙型肝炎病毒（HBV）血清免疫标志物（HBV M）和HBV DNA含量的相关性及两种指标联合检测的临床应用。方法酶联免疫吸附试验检测HBV M;荧光定量聚合酶链反应（FQ-PCR）检测HBV DNA含量。结果乙型肝炎表面抗原（HBsAg）、乙型肝炎e抗原（HBeAg）阳性的标本HBV DNA阳性率高,分别为：Ⅰ组[HBsAg、HBeAg和乙型肝炎核心抗体（抗-HBc）阳性],HBV DNA阳性率98.9%（181/183）;Ⅳ组（HBsAg和HBeAg阳性）,HBV DNA阳性率96.3%（26/27）;Ⅶ组[HBsAg、HBeAg、乙型肝炎e抗体（抗-HBe）和抗-HBc阳性],HBV DNA阳性率100.0%（2/2）;HBsAg阳性、HBeAg阴性,HBV DNA阳性率也较高,为Ⅱ组（HBsAg、抗-HBe、抗-HBc阳性）60.4%（58/96）;Ⅲ组（HBsAg、抗-HBc阳性）50%（10/20）;HBeAg、HBsAg阴性有一定的HBVDNA阳性率,但阳性率低,分别为：Ⅴ组（抗-HBs、抗-HBe、抗-HBc阳性）13.3%（2/15）;Ⅵ组（抗-HBs、抗-HBc阳性）8.3%（1/12）;Ⅷ组（抗-HBs阳性）0（0/26）;Ⅸ组（HBV M全阴性）6.7%（1/15）。结论血清HBV DNA水平与HBV M表现模式有关,与HBsAg,HBeAg有良好的相关性,而HBV M阴性的患者也可能有HBV DNA阳性,因此HBV M和HBV DNA联合检测,可有效提高乙型肝炎的检出率,为临床提供HBV感染、复制及传染性判断以及指导治疗的实验室依据。     

全血乙型肝炎病毒定量检测方法的初步研究

目的探讨聚合酶链反应(PCR)检测全血中乙型肝炎病毒(HBV)DNA含量及临床应用价值.方法取20μl血清裂解液提取HBV DNA、和取同样量的全血经不同裂解液提取血液中血清及白细胞内总HBV DNA,同时经PCR检测.对78份乙型肝炎表面抗原(HBsAg)阳性患者两种取材方法检测HBV DNA含量进行比较.结果检测HBV DNA的最低检测限为1×103拷贝/ml,29份乙型肝炎表面抗原(HBsAg)、乙型肝炎e抗原(HBeAg)和抗乙型肝炎核心抗体(抗HBc)均为阳性患者全血检测HBV DNA阳性率为100%,血清阳性率为100%,P＞0.05;37份HBsAg、抗乙型肝炎e抗体(抗HBe)和抗HBc均为阳性患者全血检测HBV DNA阳性率为97.30%,血清阳性率为48.65%,P＜0.01;12份HBsAg和抗HBc均为阳性患者全血检测HBV DNA阳性率为91.67%,血清阳性率为25%,P＜0.01;20份非乙型肝炎患者血清均为阴性.血清HBVDNA阳性多数是ALT升高患者,其拷贝数均小于全血测定的拷贝数.结论全血检测HBV DNA阳性率高,避免血液中白细胞内HBV漏检,它能准确地反应血液HBV含量,更好地为临床治疗效果作监测.     

乙型肝炎表面抗原阳性肝硬化患者前S1抗原、e抗原及乙型肝炎病毒DNA结果分析

目的探讨乙型肝炎病毒(HBV)表面抗原(HBsAg)阳性肝硬化患者血清中HBV前S1抗原(前S1抗原)、HBV e抗原(HBeAg)及HBV核酸定量检测(HBV DNA)相关性。方法 2008年7月-2011年5月对97例HBsAg阳性肝硬化住院患者和50份HBsAg阴性的健康体检者血清进行前S1抗原、HBV血清标志物检测及实时荧光定量PCR检测HBV DNA结果进行分析。结果 97份HBsAg阳性肝硬化患者血清中,前S1抗原、HBeAg及HBV DNA阳性率分别为53.6%(52/97)、22.7%(22/97)及61.8%(60/97)。22例HBeAg阳性血清中,前S1抗原阳性18例(81.8%),HBV DNA阳性20例(90.9%)。75例HBeAg阴性血清中,前S1抗原阳性34例(45.3%),HBV DNA阳性40例(53.3%),两者的前S1抗原与HBV DNA结果间都具有很好的相关性。HBV DNA含量与前S1抗原及HBeAg阳性结果显示:HBsAg阳性的肝硬化患者血清中HBV DNA阴性率为38.1%(含量<103 copies/mL),而阳性检出率HBV DNA含量主要集中在103～105 copies/mL,占81.7%(49/60),HBV DNA含量>105 copies/mL占18.3%(11/60)。结论 HBsAg阳性的肝硬化患者血清中主要以HBV非HBeAg阳性血清学模式为主,HBV DNA阳性检出率的含量主要集中在103～105 copies/mL。前S1抗原在HBeAg阳性血清中与其含有HBsAg病毒及HBeAg阳性患者具有很好的相关性,而在HBeAg阴性血清中存在着差异。     

抗HCV与血清HBV-M电化学发光检测结果相关性分析

目的回顾性分析患者抗HCV与HBV-M检出情况,探讨HCV与HBV感染之间的相互关系。方法使用电化学发光法(ECL)比较分析196例抗HCV阳性患者与32026例抗HCV阴性患者血清HBV-M单项检出率和不同模式的检出率。结果抗HCV阳性组HBV-M检出率为73.47%;抗HCV阳性组HBsAb单阳性模式检出率低于抗HCV阴性组(P<0.05);而HBcAb单阳性模式、HBeAb和HBcAb双阳性模式、HBsAg和HBcAb双阳性模式检出率却高于抗HCV阴性组(P<0.05);抗HCV阳性组单项HBsAg、HBeAg以及HbsAb的检出率与抗HCV阴性组间差异无统计学意义(P>0.05),而单项HBeAb和HBcAb阳性率却高于HCV抗体阴性组(P<0.05)。结论 HCV与HBV共感染在人群中分布面较广,共感染后存在互相干扰抑制作用,主要表现为HBcAb、HBeAb的产生,及对保护性HBsAb产生过程的干扰与抑制。     

HBV前S1抗原与HBV DNA联合检测的临床意义

目的探讨乙型肝炎病毒前S1抗原(HBV pre-Sl)、HBV DNA与HBV表面标志物(HBV-M)的关系。方法从临床乙型肝炎标本中筛出HBsAg阳性病例329例,采用ELISA法检测乙肝血清标志物和前S1抗原,荧光定量PCR法检测标本HBV DNA。结果血清HBeAg抗原阳性组,HBV DNA和pre-Sl的阳性率分别为97.2%和90.0%,而HBeAg抗原阴性组的HBV DNA与pre-Sl阳性率分别为42.9%和37.0%,HBeAg阳性患者血清的HBV DNA与pre-Sl的阳性检出率明显高于HBeAg阴性患者。结论 HBV pre-Sl和HBV DNA在各组中阳性检出率有较高的一致性,pre-Sl在一定程度上可替代HBV DNA。pre-Sl与乙肝血清标志物联合检测能为乙肝患者病毒复制、肝功能损伤提供有价值的实验室依据,同时有助于慢性乙肝患者疗效考核和预后判断。     

乙型肝炎患者胃液与血清中乙型肝炎病毒、胃部病变及传播途径的关系

目的　探讨乙型肝炎患者胃液乙肝病毒与胃部病变、血清乙型肝炎病毒 (HBV)指标及其传播途径的关系。方法　采用酶联免疫吸附试验 (ELISA)检测 5 8例慢性乙型肝炎患者及 18例血清HBV指标阴性的非乙型肝炎患者的胃液和血清中的HBV指标。用聚合酶链反应 (PCR)方法检测 5 8例中的 32例血清和胃液的HBV DNA。结果　 5 8例慢性乙型肝炎患者胃液中乙型肝炎e抗原 (HBeAg)和乙型肝炎表面抗原 (HBsAg)以及HBV DNA阳性患者其胃部病变较重 ,主要表现为隆起或平坦糜烂性胃炎、门脉高压性胃病、慢性胃炎伴胃窦急性炎症。胃液中HBeAg和HBsAg的阳性率在慢性乙型肝炎临床类型中的顺序依次为慢性乙型肝炎重度、乙型肝炎后肝硬化、慢性乙型肝炎中度 ,但HBeAg的阳性率在这 3种疾病中差异无统计学意义 (P >0 .0 16 7) ,而HBsAg的阳性率中慢性乙型肝炎重度明显高于慢性乙型肝炎中度 (P <0 .0 16 7)。 5 8例慢性乙型肝炎患者胃液中HBsAg、HBeAg及HBV DNA的检出率分别为 5 1.7%、12 .1%和 34.4 %。其胃液中的HBV指标与血清中的HBV指标无相关性 (P >0 .0 5 ) ,部分血清HBsAg、HBeAg及HBV DNA阴性者 ,胃液中出现阳性。结论　慢性乙型肝炎患者胃液中可检出HBV标志物 (HBVM )和HBV DNA ,具有传染性 ,且常伴有较明显的胃部病变 ,故应     

乙型肝炎病毒标志物 前S1抗原与HBV DNA的检测结果分析

目的探讨乙型肝炎病毒（HBV）前S1抗原（Pre-S1）与HBV血清标志物（乙肝两对半）和HBV DNA之间的关系,并分析联合检测的临床应用价值。方法收集17 145例患者血清标本并进行检测,乙肝两对半和Pre-S1抗原采用酶联免疫法,HBV DNA采用荧光定量法,对测定结果进行统计学分析。结果 HBV表面抗原（HB-sAg）、e抗原（HBeAg）和核心抗体（抗-HBc）阳性组Pre-S1抗原和HBV DNA的阳性率分别为89.2%和95.0%,而HBsAg阳性、HBeAg阴性组的Pre-S1抗原与HBV DNA阳性率分别为35.9%和43.0%,HBeAg阳性患者血清的HBV DNA与Pre-S1的阳性检出率明显高于HBeAg阴性患者。结论 HBV Pre-S1和HBV DNA在各组中阳性检出率较一致,且HBeAg阳性患者血清的HBV DNA与Pre-S1的阳性检出率明显高于HBeAg阴性患者。     

天津市某三甲医院维持性血液透析患者血源性病毒感染情况分析

【摘要】目的了解血液透析患者乙型肝炎病毒（hepatitisBvires,HBV）、丙型肝炎病毒（hepatitisCvirus,HCV）、人类免疫缺陷病毒humanimmunodeficiencyvirus,HIV）和梅毒螺旋体（treponemapal—lidum,TP）的感染情况。方法应用酶联免疫法检测449例维持性血液透析患者血清中HBV五项、抗-HCV、抗-HIV和TP抗体．应用PCR-荧光探针法检测HBsAg阳性的透析患者血清中HBV—DNA含量及抗-HCV阳性的透析患者血清中HCV—RNA含量,并进行统计学分析。结果449例透析患者HB．sAg、HBsAb、抗-HCV、TP抗体的阳性率分别为2．90％、20．71％、8．69％、1．78％,抗-HIV检测结果均为阴性。其中,HBsAg阳性患者中,HBV-DNA阳性率为46．15％,DNA拷贝数为1．1×104—3．7×10^7 IU／mL;抗-HCV阳性者中,HCV-RNA阳性率为58．97％,RNA拷贝数为8．1×10^3～1．5×10^7IU／mL。≥60岁和〈60岁两组患者HBsAg、抗-HCV阳性率比较,差异均无统计学意义（P均〉0．05）。结论血液透析患者HBV、HCV感染率相对国外较低,但感染现象在各年龄段普遍存在。TP感染情况与国内外报道相似,应加强管理防护。     

献血者TT病毒DNA及其IgG抗体的检测

目的　观察献血者输血传播病毒 (TTV)的感染情况。方法　应用 Nested- PCR对 388例献血者、16 8例非肝炎住院患者进行 TTV DNA检测 ,同时用 EL ISA法检测抗 TTV Ig G。结果　 TTV基因检出率分别为献血组 15 .46 % ,对照组 2 4.40 % ;5 5 6例受检血清中 TTV DNA总的阳性检出率为 18.17% ,抗 TTV Ig G检出率分别为献血组 13.14% ,对照组 2 7.98% ;献血组与对照组相比 TTV DNA及抗 TTV Ig G均存在显著性差异 (P<0 .0 5 )。结论　献血者存在TTV感染 ,TTV存在健康携带状态。     

Prevalence of TT virus in serum and peripheral mononuclear cells from a CAPD population.

BACKGROUND: A novel virus named TT virus (TTV) has been isolated recently from patients with posttransfusional hepatitis of unknown etiology. The prevalence of TTV in several groups at risk has been reported, however, there is no information about the prevalence of TTV in patients on continuous ambulatory peritoneal dialysis (CAPD) without blood transfusions or hemodialysis antecedents. OBJECTIVE: To study the incidence of TTV in serum and peripheral blood mononuclear cells (PBMC) of CAPD patients. DESIGN: TTV DNA was detected by polymerase chain reaction, using primers from the open reading frames (ORF) 1 and 2, in serum and PBMC from 22 CAPD patients who had not received blood transfusions or hemodialysis therapy prior to CAPD. As controls, sera from 20 patients with chronic viral hepatitis (10 with HBV and 10 with HCV) and 20 healthy donors were included in the study. RESULTS: TTV DNA was detected in the serum of 5 of 22 (22.7%) CAPD patients with both sets of primers. Four of the 5 (80%) patients with TTV DNA in their serum were TTV positive in their PBMC with primers from ORF1 and ORF2. Five of 20 (25%) patients with chronic viral hepatitis (2 patients with HBV and 3 with HCV) and 4 of 20 (20%) healthy donors were TTV DNA positive in serum. No relation was found between TTV infection and the underlying kidney disease, previous surgery, and abnormal alanine aminotransferase levels. CONCLUSION: We have found a relatively high prevalence of TTV that is similar to that found in healthy donors and in patients with chronic viral hepatitis.     

Detection of TT virus in hemodialysis patients]

Recently a newly discovered DNA virus, transfusion transmitted virus (TTV), was introduced as a cause of post-transfusion hepatitis. We studied the frequency of TTV viremia in 60 hemodialysis patients in Yamaguchi, Japan. TTV DNA was detected by heminested PCR, using primers described by Okamoto et al. TTV DNA was detected in 18 patients (30%). There was no differences in clinical characteristics, including age, gender, history of blood transfusion, and double infection of other hepatitis viruses, between TTV DNA positive patients and negative patients. Also the frequency of TT viremia was not associated with the duration of hemodialysis. These results suggest that the routes of TTV infection may be different from those of infection by HBV, HCV, or HGV.     

The clinical features of chronic hepatitis C are not affected by the coexistence of hepatitis B virus DNA in patients negative for hepatitis B surface antigen

Hepatitis B virus (HBV) DNA has been detected in the sera of hepatitis patients who are negative for hepatitis B surface antigen (HBsAg) by polymerase chain reaction (PCR). The purpose of the present study was to clarify the clinical characteristics of patients with chronic hepatitis C who are negative for serum HBsAg and positive for HBV DNA. The subjects included 49 patients with chronic hepatitis C who were negative for serum HBsAg and 119 blood donors who served as healthy controls. Serum samples were tested for the presence of HBV DNA by the nested PCR method. Serum HBV DNA was detected in 18 (37.7%) of the 49 chronic hepatitis C patients and in none (0%) of the 119 blood donors. Among the hepatitis C patients, HBV DNA was detected in 20.7% of those who were negative for all HBV-associated markers and in 57.1% of those who were positive for one or more HBV-associated marker. The HBV DNA-positive rate among those in each F stage did not significantly differ. The liver function parameters of the HBV DNA-positive and the HBV DNA-negative chronic hepatitis C patients did not significantly differ. These results suggest that hepatitis C virus is frequently coinfected with serum HBsAg-negative HBV, and that the incidence of HBV infection in blood donors is low. However, it is considered that HBsAg-negative HBV infection does not modify the blood biochemical features of chronic hepatitis C.     

Serum HBV-DNA (hepatitis B virus DNA) in acute and chronic hepatitis B infection

HBV DNA was measured in the sera of 69 patients with hepatitis B virus infections. Sixteen patients had acute hepatitis B, 24 had chronic active hepatitis (CAH), 6 had chronic persistent hepatitis (CPH), 5 had cirrhosis without CAH and 18 were asymptomatic HBsAg carriers. In patients with acute hepatitis B who recovered, HBV DNA was present in the serum transiently early in the illness. HBV DNA persisted in the serum in the two patients who developed chronic hepatitis. Sera of 23 of 24 patients with CAH were persistently positive for HBV DNA. There was no relationship between the quantity of HBV DNA in the serum and the histological intensity of activity. Thirteen of the 24 patients with CAH had histological evidence of cirrhosis in addition to CAH and HBV DNA was detected in the sera of all 13. The sera of 2 of 6 patients with CPH were positive for HBV DNA. In one it was positive only where there was clinical evidence of reactivation of HBV infection. The other patient subsequently developed CAH. Sera of 5 patients with established HBsAg positive cirrhosis but without evidence of CAH were negative for HBV DNA. Two of these patients had hepatocellular carcinoma. Sera of 18 asymptomatic anti-HBe positive carriers with normal ALT were negative for HBV DNA. HBeAg and HBV DNA were not always found in the serum together. In acute hepatitis 5 patients with HBV DNA in the serum were HBeAg positive, but in 6 patients the sera were HBeAg positive inthe absenceof HBV DNA.     

Hepatitis B virus DNA in blood donors with anti-HBc as a possible indicator of active hepatitis B virus infection in Yucatan, Mexico

Hepatitis B virus (HBV) may be present in serum even when negative for HBV surface antigen (HBsAg). If routine screening of sera for anti-HBV core antigen (anti-HBc) is not done, low-level HBV viraemia may not be identified. A study was done on the presence of HBV DNA in serum samples from Mexican blood donors negative for HBsAg. Sera from 158 volunteer blood donors, negative for HBsAg and anti-HBs, but positive for anti-HBc, were analysed using nested polymerase chain reaction (PCR). HBV DNA was detected in sera from 13 (8.23%) of the 158. Specificity of the PCR-amplified products was corroborated using Southern blot. Single strand conformation polymorphism (SSCP) analysis showed identical SSCP-banding patterns for all 13 PCR products, suggesting similar cDNA sequences. Occult HBV infection was observed in approximately 8% of anti-HBc only donors. The absence of HBsAg in the blood of apparently healthy individuals may not be sufficient to ensure lack of circulating HBV, and blood containing anti-HBc only may be infectious until proven otherwise.     

免疫筛查阴性献血者血样病毒核酸检测的研究

目的了解二次酶联免疫筛查献血者血样漏检的原因。方法将二次酶联免疫筛查阴性的献血者血样在加样仪上实现血液样本汇集,用全自动核酸提取仪提取样本核酸,以核酸扩增检测仪做HBV、HCV和HIV自动扩增检测。对HBsAg阴性、HBVDNA阳性献血者用核酸筛查试剂定量检测HBV,并每隔2周对其跟踪采血,做HBV两对半免疫检测和HBsAgV3的确认试验。结果16320份二次酶联免疫筛查阴性的合格献血者血样中,8份HBVDNA阳性(漏检率0.49‰),未发现HCV和HIV1RNA阳性。8份HBVDNA阳性献血者乙肝两对半免疫检测HBsAg、HBsAb和HBeAg均为阴性,HBcAb均为阳性,3份HBeAb为阳性。6例HBsAg阴性HBVDNA阳性献血者血样的病毒滴度在(76～1490)copies/ml,2例病毒滴度过低,未定量检测到病毒。跟踪6名HBVDNA阳性献血者,1例18周时HBsAg确认试验阳性,其余5例仍为阴性。结论现行的二次酶联免疫技术的血液筛查存在HBV漏检,原因可能是隐匿性乙型肝炎病毒感染。应重视血液筛查工作中HBV的漏检及输血传播,并在现有的血液筛查模式中或增加HBcAb检测,或增加病毒核酸筛查。     

二氧化硅提取核酸用于多聚酶链反应

利用硫氰酸胍裂解病毒和二氧化硅颗粒能够特异性吸附核酸的特性，将二氧化硅悬液、裂解液及肝炎病毒感染血清共同保温，病毒裂解释放出的核酸结合在二氧化硅颗粒上，经过双蒸水洗脱，所得核酸直接作ＰＣＲ扩增。     

2260例静脉注射海洛因依赖者HBV、HCV感染状况分析

应用ELISA法对 2260例iv海洛因依赖者进行血清HBsAg、抗 -HCV、ALT检测。结果表明 ,iv海洛因依赖者血清HBsAg、抗 -HCV阳性率分别为 37.8%、53.9% ,HBsAg、抗 -HCV同时阳性者 4 56例 ,重叠感染率 20.2 %均显著高于对照组 (24.9%、22.3%、10.8% )。两组之间的差异有非常显著性 (均P 0.05)。在iv海洛因依赖者中 ,HCV感染率较HBV感染率更高 ,抗 -HCV阳性者 ,ALT异常率高于HBsAg阳性者。提示iv毒品传播HCV已成为一种不可忽视的重要途径。     

Prevalence and occurrence of infections caused by hepatitis B virus in the dialysis units in the Apulia region

Sera of 803 hemodialysis patients and 413 staff members were tested to evaluate the relationship between infectivity markers and spread of HBV infection in dialysis units. HBsAg was detected in 13.8% patients undergoing chronic hemodialysis and 3.9% staff members. High prevalence of HBeAg and DNA polymerase activity was observed only in HBsAg positive patients. The highest titers of HBsAg and anti-HBc were detected in hemodialysis patients, whereas asymptomatic carriers showed low titers of these markers. A highly significant correlation was recorded between detection of HBeAg in patients and presence of serum DNA activity. These data suggest that in HBsAg hemodialysis patients a more active viral replication occurs and a higher contagiousness of these subjects.