丙型肝炎患者的TT病毒感染状况分析

[目的 ]了解延边地区丙型肝炎患者感染TT病毒的状况 .[方法 ]采用ELISA法检测 5 7例丙型肝炎患者的TT病毒 IgG ;采用PCR技术检测TT病毒DNA .[结果 ]TT病毒 IgG的阳性率为 37%(2 1/ 5 7) ,TT病毒DNA的阳性率为 4 2 % (2 4 / 5 7) .[结论 ]延边地区丙型肝炎患者中存在TT病毒的重叠感染 .     

Relationship of hepatitis C virus infection with primary hepatic carcinoma: an investigation of serum HCV RNA in different population groups

A preliminary study on the serum hepatitis C virus(HCV)RNA in 182 pa-tients with different chronic liver diseases and 35 blood donors was carried out with“nested”polymerase chain reaction.It was found that serum HCV RNA was detected in36.6%(34/93)cases of primary hepatic carcinoma(PHC),31.7%(20/63)liver cirrhosis,15.4%(4/26)chronic hepatitis and 2.9%(1/35)blood donors;most of the HCV RNA pos-itive cases(41/58)were complicated with HBV replicating markers;the rate of single posi-tive for HCV RNA was 15.1%(14/93)in PHC,3.2%(2/63)in liver cirrhosis,and 3.8%(1/26)in chronic hepatitis.These findings imply that about 20% of PHC cases appear tobe related to the coinfection of HCV and HBV and 15% of PHC cases are related to sin-gle HCV infection.     

微孔板固相杂交法检测丙型肝炎病毒

目的 寻找一种快速、准确、简便的丙型肝炎病毒（hepatitis C virus，HCV）RNA逆转录-聚合酶链反应产物的检测方法。方法 所有标本均作逆转录-套式聚合酶链反应，其产物分别以微孔板固相杂交法及聚丙烯酰胺凝胶电泳法检测，对结果可凝及阴性者均以HCV Amplification KIT对其血清再作扩增并检测，并以此为金标准进行比较。结果 144份血清（其中抗-HCV阳性64例），微孔板固相杂交法阳性70例，灵敏度97.18%，特异度98.63%，聚丙烯酰胺凝胶电泳法阳性61例，灵敏度81.69%，特异度95.89%，取同一份产物倍比长稀释后再作检测，前者可检测出2^-9稀释度，后者检测出2^-7稀释度。结论 微孔板固相杂交法检测丙型肝炎病毒快速、准确、简便，并且可定量，尤其适用于大批量标本的检测，但于基层推广。     

Hepatitis C virus infection in different groups of children in Wuhan area

Summary To investigate the incidence of child’s HCV infection in our area, 637 children with different background, including 65 posttransfusion cases, 419 hepatitis patients (250 cases of acute hepatitis A, 156 cases of chronic hepatitis B and 13 cases of non-A, non-B hepatitis), 50 infantile hepatitis syndrome (1HS) infants and 103 healthy day-cared children were tested for serum anti-HCV antibody (EIA) and HCV RNA (nested PCR). It was found that posttransfusion children had significantly higher anti-HCV positive rate (30. 8%) and HCV infection incidence (43.1%) than hepatitis patients (4.3% and 5.3%), IHS infants (6.0% and 8.0%) and daycared children (2.9% and 2.9%). 25 of 33 cases with posttransfusion hepatitis (PTH) developed hepatitis C, which was the leading cause of PTH (75.8%) and NANB PTH (25/30, 83.3%). The incidence of HCV infection in NANBH patients was 23.1% (3/13) which was apparently higher than that in day-cared children (P <0. 02) and lower than that in PTH patients (P<0. 001), but not statistically different from that in AHA and CHB patients (P>0. 05). Mother-infant paired study in IHS group showed that 4 pairs of mother-infant had HCV infection, one boy aged 8 months and his mother were anti-HCV positive, and another 3 pairs possessed HCV RNA in sera. 3 of 103 healthy day-cared children were found to have inapparent HCV infection, who were anti-HCV and HCV RNA positive.     

丙型肝炎病毒NS5B区基因的扩增、克隆及序列分析

目的 对丙型肝炎病毒(HCV)NSSB区基因克隆并测序，研究其变异状况。方法 从HCV RNA阳性血清中抽提病毒RNA，利用巢式RT-PCR对NS5B区基因进行扩增，将扩增获得的靶cDNA克隆至P^KPL-3a质粒载体中，并以内引物为测序引物进行序列测定分析。结果 构建了已克隆HCVNS5B区基因的重组质粒KHC5—1；序列分析表明，与HCV-BDS株同源性最高，属于主要流行于我国的HCV 1b基因亚型。结论 证实了在NS5B区，与RNA依赖的RNA聚合酶(RDRP)活性密切相关的基元结构Gly-Asp-Asp及其相邻序列的同源性非常高。     

External quality assessment on detection of hepatitis C virus RNA in clinical laboratories of China

Background As with many studies carried out in European countries, a quality assurance program has been established by the National Center for Clinical Laboratories in China （NCCL）. The results showed that the external quality assessment significantly improves laboratory performance for quantitative evaluation of hepatitis C virus （HCV） RNA.
Methods Serum panels were delivered twice annually to the clinical laboratories which performed HCV RNA detection in China. Each panel made up of 5 coded samples. All laboratories were requested to carry out the detection within the required time period and report on testing results which contained qualitative and/or quantitative test findings, reagents used and relevant information about apparatus. All the positive samples were calibrated against the first International Standard for HCV RNA in a collaborative study and the range of comparison target value （TG） designated as ±0.5 log.
Results The numbers of laboratories reporting on qualitative testing results for the first and second time external quality assessment were 168 and 167 in the year of 2003 and increased to 209 and 233 in 2007; the numbers of laboratories reporting on quantitative testing results were 134 and 147 in 2003 and rose to 340 and 339 in 2007. Deviation between the mean value for quantitative results at home in 2003 and the target value was above 0.5 log, which was comparatively high. By 2007, the target value was close to the national average except for the low concentrated specimens （10^3 IU/ml）. The percentage of results within the range of GM±0.5 log10 varied from 8.2% to 93.5%. Some laboratories had some difficulties in the exact quantification of the lowest （3.00 log IU/ml） as well as of the highest viral levels （6.37 log IU/ml） values, very near to the limits of the dynamic range of the assays.
Conclusions The comparison of these results with the previous study confirms that a regular participation in external quality assessment （EQA） assures the achievement of     

慢性乙型肝炎患者196例病毒基因分型与临床的关系

目的探讨西安市中心医院感染科诊治的慢性乙肝患者血清中HBV基因型的分布状况及HBV基因型与临床特征的关系。方法采用巢式PCR法对196例慢性HBV患者的血清进行病毒基因型检测,并分析HBV基因型与患者临床特征的关系。结果 196例慢性HBV患者的基因分型结果为C型138例(70.4%)、B型40例(20.4%)、B/C混合型18例(9.2%),基因型的分布在性别上无统计学差异;各基因型在无症状携带者、慢性乙肝、肝硬化、肝癌中的分布差异也无统计学意义(P>0.05);C型感染者血清HBeAg阳性率(75.2%)高于B型感染者(14.2%),差异有统计学意义(P<0.05)。各基因型患者在HBV-DNA载量和ALT水平上无统计学差异(P>0.05)。结论西安市中心医院感染科诊治的HBV患者的基因型以C型和B型为主;基因C型患者血清HBeAg阳性率高于B型;基因C型患者与B型相比在肝脏炎症程度、HBV-DNA载量方面无显著差异;基因B型和C型患者临床特点相似。     

Detection of hepatitis C virus in serum and peripheral blood mononuclear cells by two reverse transcription polymerase

Over 75% of patients with acute hepatitis C （HCV） will develop into chronic infection associated with chronic hepatitis, end stage liver disease or hepatocellular carcinoma. HCV infection is usually ignored because the level of HCV in serum of some patients is very low and most patients with chronic HCV infection are asymptomatic. Some patients with persistent abnormal liver function may be infected with HCV although HCV RNA and HCV antibodies in serum are negative because their HCV load is under the current detectable limit or sometimes HCV is absent in serum but present in the liver or peripheral blood mononuclear cells （PBMCs） and other tissues.     

丙肝病毒感染对C反应蛋白表达的影响

目的：探讨丙型肝炎病毒(HCV)在体内外对C反应蛋白(CRP)表达的影响。方法收集武汉大学中南医院2015年1月至2016年2月HCV感染患者标本106例和健康体检者标本110例,免疫透射比浊法检测其血清CRP水平,采用逆转录聚合酶链反应(RT-PCR)检测HCV感染的Huh7.5.1细胞及其对照细胞中CRP mRNA的表达。结果 HCV感染患者血清CRP水平为(8.62±5.56) mg/L,显著高于健康体检者的(1.35±0.71) mg/L,差异有显著统计学意义(P<0.01);HCV感染的Huh7.5.1细胞中CRP mRNA的表达水平较对照细胞高,其CRP/β-actin灰度比值分别为0.31和1.5。结论 HCV感染能够上调肝细胞CRP的表达。     

丙型肝炎病毒母婴传播的研究

目的:研究丙型肝炎病毒(HCV)母婴传播机率及感染方式.方法:用ELISA和RT-PCR法检测母婴血清及母亲乳汁、羊水、唾液的抗-HCV IgG及HCV-RNA,循环测序法测定母婴HCV-cDNA序列.结果:对1 521例孕妇筛查,其中15例抗-HCV IgG阳性(12例HCV-RNA阳性).所生15例婴儿,随访结果仅1例抗-HCV IgG持续阳性,出生时2例血清HCV-RNA阳性者中的1例抗-HCV IgG及HCV-RNA持续阳性.HCV母婴传播率为16.7%(2/12).其中一对母婴间HCV-cDNA序列同源性为100%.羊水、乳汁、唾液抗-HCV及HCV-RNA检出率分别为100%和25.0%,53.5%和16.7%,100%和0.0%.结论:HCV母婴传播可能发生于宫内或分娩时,羊水可能是引起HCV母婴传播的重要因素,乳汁、唾液引起传播可能性较小.     

丙肝病毒核心抗原检测在丙肝病毒感染诊断中的作用研究

目的探讨丙肝核心抗原检测在丙肝感染诊断中的作用。方法收集175例丙肝抗体检测标本同时检测HCVAb、HCV游离抗原、HCV总抗原、HCVRNA。结果 75例HCVAb阳性反应标本,检出HCV游离抗原阳性17例,HCV总抗原阳性54例,HCVRNA阳性50例;100例HCVAb阴性反应标本,检出HCV游离抗原阳性1例,HCV总抗原阳性2例,HCVRNA阳性1例。结论 HCV核心抗原是HCV早期感染的标志之一,检测HCV核心抗原有利于HCV感染的早期诊断。     

HBV-DNA的含量与宫内感染的关联性

目的：探讨孕妇血清中HBV—DNA含量与胎儿宫内感染及造成宫内感染的相关因素．方法：用ELISA方法筛选HBsAg阳性孕妇228例,并用定量PCR技术检测HBsAg阳性孕妇血清以及脐血中HBV—DNA．新生儿根据有无HBV感染分为胎儿感染组及非感染组（对照组）,调查宫内感染的相关因素,结果：HBsAg,HBeAg、抗HBc阳性孕妇的新生儿脐血HBV—DNA捡出率21％（14／68）;HBsAg．抗HBe,抗HBc阳性者为1．7％（2／117）;HBsAg和HBeAg双阳性者为20％（3／15）;HBsAg和抗HBc阳性者为11％（2／19）;HBsAg单一抗原阳性者脐血中未捡出HBV—DNA．胎儿总感染率为9,2％（21／228）．胎儿宫内感染率随孕妇血清中HBV—DNA含量增加而升高,宫内感染的危险性越大．胎儿感染组与非感染组在多种临床相关因素中先兆早产孕妇胎儿感染者多于非感染组（P〈0．05）．结论：孕妇不同的感染状态影响宫内HBV感染率,HBeAg与HBV—DNA高滴度是胎儿宫内感染的高危因素．孕妇出现先兆早产使胎儿的感染率增加．     

丙型肝炎病毒核酸单引物聚合酶链反应产物的直接序列分析

利用反转录聚合酶链反应(RT-PCR)技术,得到丙型肝炎病毒核酸非编码区引物所引导的扩增片段。用改进的不对称PCR方法,制备单链cDNA,直接作为模板进行序列分析,结果与Takamizawa发表的序列符合。本法简单方便,亦可用于其他类似的研究中。     

丙型肝炎病毒感染与原发性肝癌的关系——不同人群血清HCV RNA的调查

本文以聚合酶链反应(PCR)对重庆地区各种慢性肝病(CLD,n=182)和献血员(n=35)血清HCV RNA的存在状况进行了初步调查。结果发现:①HCV RNA在原发性肝癌(PHC)、肝硬化(LC)、慢性肝炎(CH)及献血员中的检出率分别为36.6％(34/93)、31.7％(20/63)、15.4％(4/26)和2.9％(1/35);②多数HCV RNA阳性CLD(41/58)与HBsAg或HBV DNA双阳性,单纯HCV RNA阳性见于15.1％(14/93)PHC、3.2％(2/63)LC和3.8％(1/26)CH。此资料提示约20％PHC可能与HCV-HBV双重感染有关,约15％PHC可能与单独HCV感染有关。     

慢性肝病与乙型、丙型肝炎病毒感染的关系

采用酶联免疫法（ELISA）对270例慢性肝病和对照组103例非肝病患者进行了HBV标志物及抗－HCV的检测。结果显示；慢性肝病HBV感染率为85．2％，抗－HCV阳性率为23．7％，HBV和HCV双重感染率为17．8％，单纯抗－HCV阳性率为5．9％。对照组HBV感染率为14．6％，抗－HCV阳性率为5．8％。126例有输血史的慢性肝病患者抗－HCV阳性率（42．1％）明显高于144倒无输血史者（7．6％），两组差异有显著性（P＜0．01）。提示HBV感染是引起慢性肝病的主要病因，HCV次之。HCV感染与输血有密切关系。     

胎儿宫内感染乙型肝炎病毒的实验研究

目的探讨胎儿宫内乙型肝炎病毒(HBV DNA)感染的实验诊断方法及其临床意义.方法应用酶联免疫吸附试验(ELISA)和荧光定量聚合酶链反应(FQ-PCR)对156例乙肝表面抗原(HBsAg)阳性孕妇的母血和产后婴儿脐血及婴儿外周血的乙型肝炎血清学标志物(HBV-M)和HBV DNA进行检测.结果156例HDsag阳性孕妇所生婴儿脐血和婴儿外周血HBsAg的检出率为8 3%(13/156)、6.4(10/156),HBeAg检出率分别为7.1%(11/156)、5.1%(8/156),母血、脐血、婴儿外周血HBV DNA阳性检出率分别为36.5%(57/156)、28.2%(44/156)、23.7%(37/156);57例HBV DNA阳性孕妇血病毒含量(拷贝数/ml的对数值)为7.34±2.28,37例阳性婴儿外周血HBV DNA含量为6.08±0.96;与婴儿外周血检测结果相比较,脐血中HBsag、HBeAg的阳性率不仅存在23.1%和37.5%的假阳性,HBV DNA也存在16.7%(7/42)的假阳性;在HBeAg或HBV DNA阳性孕妇中,婴儿外周血HBV DNA的检出率分别为73.8%(31/42)、64.9%(37/57),显著高于HBeAg或HBV DNA阴性者5.3%(6/114),(P＜0.01):婴儿外周血中HBV DNA阳性率随孕妇HBVDNA含量增加而显著增加(P＜0.005),其HBV DNA的含量与母血呈正相关,(r=0.39).结论婴儿脐血HBV检测是诊断胎儿宫内HBV感染的筛选指标,而婴儿外周血的检测才具有确诊意义;HBV DNA定量检测是胎儿宫内HBV感染和感染程度最为直接、敏感的诊断方法;孕妇HBeAg、HBV DNA阳性是胎儿宫内感染HBV的高危因素.     

乙型肝炎病毒S区和C区基因长片段扩增新方法的探讨

目的 :为HBV基因研究提供一个灵敏度和扩增效率均较佳的长片段PCR新方法 .方法 :应用自行设计的引物扩增血清中HBVS区和C区的 2 .5kb基因 ,并与HBV全长扩增方法进行比较 .结果 :HBV 2 .5kbPCR方法在HBsAg阳性患者血清中的阳性检出率明显高于全长PCR方法 (P <0 .0 5 ) ,相同反应条件下 2 .5kbPCR产物的平均A值和灰度总值也高于 3.2kb的全长基因组 (P <0 .0 5 ) .结论 :HBV2 .5kbPCR方法的灵敏度和扩增效率高于全长PCR方法 ,可作为HBV相关疾病研究中的一种新的技术手段     

湖南地区输血传播肝炎病毒感染者的初步观察

为观察湖南地区输血传播病毒（ＴＴＶ）的感染情况,合成了特异性引物,采用巢式聚合酶链反应两次扩增血清ＴＴＶＤＮＡ。共检测７０例肝炎病人血清和４３例献血员血清。结果显示：在３８例血清ＨＢｓＡｇ阳性、ＨＢｅＡｇ和ＨＢＶＤＮＡ阴性的病人中,１０例ＴＴＶＤＮＡ阳性（２６３％）,ＡＬＴ平均为（４５２±２３６）Ｕ·Ｌ－１;３２例血清甲～戊型和庚型肝炎标志物阴性的病人中,１２例ＴＴＶＤＮＡ阳性（３７５％）,ＡＬＴ平均为（５８２±２５９）Ｕ·Ｌ－１;４３例献血员中,４例ＴＴＶＤＮＡ阳性（９３％）,ＡＬＴ在正常范围。提示湖南地区首次发现ＴＴＶ感染者。本资料与日本首次报道者相近;不能排除ＴＴＶ感染与ＡＬＴ升高的关系。