丙型肝炎病毒核酸定量检测的临床意义

目的　研究丙型肝炎患者血清中的丙型肝炎病毒核糖核酸 (HCVRNA)载量与抗丙型肝炎病毒抗体 (抗 HCV)之间的相关性。方法　用荧光定量 (FQ PCR)法检测 2 0 8例疑诊丙型肝炎患者血清中的HCVRNA ,酶联免疫吸附法(ELISA)检测抗 HCV。结果　 2 0 8例血清样本中HCVRNA阳性率为 81.3 %。HCVRNA平均载量为 4.63× 10 5Copies/ml。结论　FQ PCR法检测HCVRNA是判定HCV感染的直接证据 ,是反映病毒复制与传染性的直接指标 ,有确诊价值与抗病毒治疗疗效评估价值。HCVRNA与抗 HCV具有高度正相关。HCVRNA较抗 HCV检出率有显著性差异     

丙型肝炎病毒不同功能区抗体与病毒核糖核酸的相关性

目的 探讨慢性丙型肝炎患者体内不同功能区抗体的免疫应答及与HCV RNA浓度的关系。方法 克隆表达丙型肝炎病毒不同功能区优势表位抗原蛋白(HCV—C、NS3、NS4、NS5和HVR1)，建立单片段ELISA法检测抗HCV抗体，并以定量HCV RT—PCR法检测患者血清中HCV RNA浓度。结果 慢性丙型肝炎患者体内HCV不同功能区抗体检出有较大的差异，HCV—C、HVR1检出最高，其次为NS3、NS4和NS5。结论 HCV—C、NS3、NS4、和HVR1抗体滴度与HCV RNA浓度均有较好的相关性。     

大肠杆菌高效表达的HCVE2蛋白在丙型肝炎检测中应用

为探讨大肠杆菌高效表达的HCV E2蛋白在丙型肝炎诊断中应用价值,采用了238份HCVRNA阳性血清标本进行了E2抗体和抗HCV测定,方法采用ELA,和PCR法.结果有164份E2抗体阳性,占68.9%,卡方检验P＞0.05;用抗HCV检测与E2抗体测定方法比较,两种方法无显著性区别（P＞0.05）;在HCV RNA阳性,抗HCV阴性的血清中,有半数可测出E2抗体的阳性.提示在大肠杆菌中高效表达了可溶性的HCV E2蛋白,纯化后可进行E2抗体测定,且E2抗体阳性数与HCVRNA,抗HCV阳性数无显著性差别,E2抗体可以检出一部分抗HCV检测不能检出的HCV感染者.     

体外感染实验探讨丙型肝炎病毒的ADE分子机制

目的研究丙型肝炎病毒(HCV)在感染人滋养层细胞的过程中是否存在抗体依赖性感染增强(ADE)作用,以探讨HCV母婴传播的分子机制。方法将HCV阳性血清以4种不同方式感染人滋养层细胞,以免疫电镜观察HCV病毒颗粒在滋养层细胞中的表达,应用RT-PCR法、免疫组化法检测细胞内外的HCVRNA正链、负链,HCVNS5、NS3及C区抗原。结果在滋养层细胞胞浆内发现了HCV病毒颗粒,HCVRNA正链、负链,HCVNS5、NS3、C区抗原仅在全血清感染细胞内或上清中间断测得。结论HCV可以在滋养层细胞中复制。HCV感染滋养层细胞的过程中存在ADE机制,抗体和补体均参与了HCV的跨膜转运过程。     

抗中性粒细胞胞质抗体的检出率及其靶抗原研究

目的　了解抗中性粒细胞胞质抗体 (ANCA)阳性检出率、流行病学特点及其靶抗原。方法　应用间接免疫荧光法 (IIF)、抗髓过氧化物酶 (MPO)和抗蛋白酶 3(PR3)酶联免疫吸附试验(ELISA)对近年送检的怀疑小血管炎的 5 6 0 4例患者血清进行了检测 ,对IIF ANCA阳性而抗MPO和抗PR3抗体均阴性的血清还进行了其他 5种ANCA特异性靶抗原的检测。并初步对ANCA阳性患者流行病学特点进行分析。结果　IIF ANCA检出率为 5 3% ,阳性检出最多在 7、8及 12月份。另外所有血清中有 390例 (7% )ANA阳性。所有血清进行抗MPO和抗PR3 ELISA检测 ,抗MPO抗体阳性 2 13例 ,抗PR3抗体阳性 32例 ,两者同时阳性 5例。 4 8例不识别MPO和PR3而IIF法阳性的血清中 13例识别其他已知靶抗原 ,识别杀菌 /通透性增高蛋白 (BPI)、人弹力蛋白酶 (HLE)、组蛋白酶G (CG)、天青杀素 (AZU)和乳铁蛋白 (LF)等靶抗原的血清分别为 7、5、1、1、0例 ,其中 1例为抗BPI和抗HLE ANCA同时阳性。 85 %的IIF ANCA阳性患者确诊为ANCA相关小血管炎。这些患者中 ,抗MPO和抗PR3的比例约为 7∶1:男女比例为 1∶1 12 ,年龄 7～ 79岁 ,平均 5 3 1岁 ,>6 0岁的老年人男女比例为 1 17∶1,而年龄 <2 0岁患者男女比例为 1∶4。结论　ANCA相关疾病在我国并不少见 ,以IIF法检     

应用丙型肝炎病毒合成肽检测HCV特异性IgM抗体

目的筛选验证捕捉丙型肝炎病毒(HCV)抗体能力最强的合成肽,进一步探讨抗HCV-IgM的效价与HCVRNA含量之间的关系。方法合成HCVCp14、HCVCp9和HCV5-1-1三种肽段,用酶联免疫吸附法(ELISA)测定乙丙型肝炎和抗HCV阳性者血清抗HCV-IgM。同时测定HCVRNA。结果在39例急性丙型肝炎患者中,抗HCVCp14-IgM的阳性率为89.7%,而HCVCp9-IgM和抗HCV5-1-1-IgM的阳性率分别为48.7%和17.9%。HCVCp14-IgM的效价(A492nm值)>2.0以上者,HCVRNA的平均含量为105.1±3.6copies/μl。结论抗HCVCp14-IgM的检出及效价高低与HCV的复制活跃程度相关。     

部队中丙型肝炎病毒(HCV)感染状况及其基因型的研究

本文应用ELISA、RT-PCR法。对1200名陆、海、空三军官兵、858名新兵及76名献血员开展了HCV感染的血清流行病学调查分析。陆、海、空三军官兵的抗-HCV和HCVRNA阳性率分别为2.67％及2.25％。这一结果不仅表明在部队人群中存在HCV感染,而且多为具有传染性的隐性感染者。不同籍贯新兵的抗HCV和HCVRNA的阳性率分别为1.86％及     

抗-HCV-IgG阳性血清检测HCV的临床意义

了解抗-HCV-IgG阳性时、仅肝功能轻度异常、与HCV感染者有接触史时等临床情况下HCV在血液内存的几率,验证在这些临床情况时开展检测HCV确证试验的临床意义.采集血清为检测标本,用ELISA技术检测抗-HCV-IgG,RT-PCR技术检测HCV-RNA.血清HCV-RNA的检出率,抗-HCV-IgG阳性组为41.9%、仅肝功能轻度异常组为25.7%、无症状体检组为29.4%.临床对抗-HCV-IgG阳性者、仅肝功能轻度异常者、有接触史者等特殊人群应做检测HCV的确证试验,RT-PCR技术可以用做临床检测HCV的确证试验方法.     

48例原发性肝癌患者血清丙型辟炎抗体检测

为探讨PHC与丙型肝炎的关系．采用ELlSA方法检测4B例0PHC及106例非肝病患的扰HCV抗体及HBV-M．结果显示PHC组中抗HCV抗体阳性2l例(43.8％)．对照组14例(13．2％．P＜0.01)HBV-M阳性45例(93．8％)．对照组50例(47．2％．P＜0．01)。HBV-M阳性PHC病人中．HBsA8、HBeAb及HBeAb三项阳性21例(46．7%)．HAg、HBeAg及HBcAb三项阳性7例(15．5％)，HBsAg及HBcAb二项阳性5例(11.1％)．HBV-M阴性PHC患中，抗HCV抗体阳性2／3例(66．7%)．HBV-M阳性PHc病人中．抗HCV抗体阳性19／45例(42．2%．P＜0．05)；HBsAg阳性PHC病人中，扰HCV抗体阳性14／38例(36．8%),HBsAg阴性病人中，扰HCV机体阳性7／10例(70．0％．P＜0．05)。在抗HCV抗体阳性PHC患中，3个月前有明确输血史8例(38．1%)．以上结果提示在我国，PHC与丙型肝炎有关，在HBV-M阳性患中，HBaAg、HBeAb及HBcAb兰项阳性患发生PHC可能性最大。     

慢性丙型肝炎病毒感染状态与机体免疫功能关系的探讨

目的：探讨慢性丙型肝炎病毒感染患者HCVRNA增殖状态与T细胞亚群功能及血浆中IL-2和sIL-2R活性的关系。方法：以间接免疫荧光法，ELISA法和RT-PCR分别检测75名慢性HCV感染患者外周血T细胞亚群，IL-2及sIL-2R的水平和HCVRNA。结果：慢性HCV感染患者周围血CD3 ，CD4 淋巴细胞亚群，CD4 ／CD8 比值及IL-2水平均显著低于正常对照组(P<0.01)而sIL-2R明显升高(P<0.01)；血清HCVRNA阳性患者T细胞亚群，CD4 ／CD8 比值及IL-2水平显著低于HCVRNA阴性患者(P<0.01)。结论：慢性HCV感染患者的机体免疫功能紊乱，细胞免疫功能低下，HCVRNA阳性患者较阴性患者更甚，提示细胞免疫功能受抑可能是HCV持续增殖的原因。     

Seven-years follow-up on trial of Interferon alpha in patients with HCV RNA positive chronic hepatitis C

AIM To study the long-term efficiency of therapy with Interferon alpha (IFN-a) in patients with HCVRNA positive chronic hepatitis C.METHODS Ten patients were enrolled in the study, whose age 31 -62 years (mean 53 years), course 6- 72months (mean 24 months), of whom, 6 patients with mild CHC, 4 moderate CHC. All patients receivedIFN-a 3 MU three times weekly for six to twelve months, and then followed up for seven years after the endof treatment. The results of hepatic functions and HCV RNA at the end of treatment and follow-up period inall patients were observed.RESULTS ( At the end of treatment, clinical symptoms recovered obviously in all patients, virologicalresponse (defined as HCV RNA loss) occurred in 5 of 7 (71.4%) patients (<60 years old) and in 1 of 3(33.3%) patients ( >60 years old). At the end of follow-up, the rates of HCV RNA loss were 42.9% (3/7)and 33.3% (1/3), respectively, in these group. Virological sustained response (defined as HCV RNA loss atthe end of treatment and follow-up) occurred in 3 of 6 (50%) patients (6 - 12 month-course) and in 1 of 4(25%) patients (> 12 month-course). A sustained HCV RNA response was observed in 2 of 7 (28.6%)patients with IFN-a therapy for 6m and in 2 of 3 (66.7%) patients with IFN-a therapy for more than 6 m. Ofall patients, 4 patients with sustained HCV RNA response were mild CHC, 4 patients with sustained HCVRNA positive were mild CHC (2 patients), moderate CHC (2 patients), respectively; other 2 patients withHCV RNA loss at the end of treatment but recurred at the end of follow-up, were moderate CHC. ②Biochemically sustained response (defined as ALT normalization at the end of treatment and follow-up) wasobserved in 5 out of 10 (50%) patients, and these 5 patients were mild CHC, of whom, 4 patients with HCVRNA sustained negative, 1 patient with HCV RNA loss and then recurred again. Two patients with ALTnormalization at the end of follow-up were one mild CHC, one moderate CHC, respectively. Other 3patients with no response were moderate CHC, of whom, 2 patients with HCV RNA sustained positive, 1patient with HCV RNA loss then recurred, and in these 3 patients, the lower limits of ALT were more than121 U/L- 148 U/L. ③ Of 10 patients, 3 moderate CHC patients were far from satisfactory to IFN-αtherapy, of whom, 2 coinfected with HBV, t with post-hepatitis cirrhosis.CONCLUSION The CHC patients with younger age, shorted course, and lighter liver changes in biopsy(mild CHC) have better response to IFN-α therapy, and the efficiency of therapy with IFN-α for 12 m aremore satisfactory than those for 6 m. The patients with coinfected HCV and HBV have a response to IFN-αtherapy worse than the others.     

外周血单核细胞中丙型肝炎病毒RNA正负链检测的临床意义

目的通过对外周血单核细胞（ＰＢＭＣ）中ＨＣＶＲＮＡ正负链的检测,来探讨其与丙型肝炎慢性化及干扰素治疗的关系．方法慢性丙型肝炎患者４０例,其中干扰素治疗者１０例,分离血清及ＰＢＭＣ．异硫氰酸胍－酚－氯仿抽提法提取ＨＣＶＲＮＡ,应用逆转录－巢式ＰＣＲ技术检测ＨＣＶＲＮＡ正负链．结果血清正链ＨＣＶＲＮＡ阳性率为６７５％,但负链均为阴性,ＰＢＭＣ中正链ＨＣＶＲＮＡ阳性率为５７５％,负链的阳性率为３５０％．其中３例患者血清中正链ＨＣＶＲＮＡ为阴性,而ＰＢＭＣ中为阳性．１０例干扰素治疗者在治疗结束时血清正链ＨＣＶＲＮＡ６０％转阴,ＰＢＭＣ中负链ＨＣＶＲＮＡ８０％转阴,而正链仅３７５％转阴．结论ＨＣＶ能在ＰＢＭＣ中存在和复制,这可能是导致丙型肝炎易发生慢性化的原因之一．ＰＢＭＣ中ＨＣＶＲＮＡ正负链的检测对于临床判断干扰素的疗效及预后有重要意义     

血液透析患者丙型和庚型肝炎病毒感染的研究

为了解血液透析患者中丙型肝炎病毒(HCV)和庚型肝炎病毒(HGV)的感染情况,并探讨相对危险因素,对48例在302医院和武警总医院进行维持性血液透析的患者用逆转录聚合酶链(PCR)反应和酶联免疫法检测了血清中HCV RNA、HGV RNA及其抗体水平。结果显示,抗-HCV和HCV RNA阳     

血清抗－HCV与HCV RNA检测结果比较

采用逆转录一套式－聚合酶链反应（ＲＴ－ｎｅｓｔｅ－ＰＣＲ）及ＥＬＩＳＡ（Ａｂｂｏｔｔ）对一组经第二代抗－ＨＣＶＥＬＩＳＡ（上海科华生物技术有限公司，科华）试剂筛选的抗－ＨＣＶ阳性血清及抗－ＨＣＶ阴性的非甲非乙型肝炎患者血清，进行了ＨＣＶＲＮＡ与抗－ＨＣＶ检测。对两者检测结果及相互关系进行了分析与比较。结果显示：Ａｂｂｏｔｔ与科华ＥＬＩＳＡ试剂两者抗－ＨＣＶ检测的阳性符合率及总符合率为７７．７％及７９．８％。两者与ＨＣＶＲＮＡ检测阳性结果的符合率分别为４９，７％及５６．３％。２０８份血清中ＨＣＶＲＮＡ阳性１０９份，其中抗－ＨＣＶ阳性组ＨＣＶＲＮＡ阳性率为６０．２％，较之抗－ＨＣＶ阴性之非甲非乙型肝炎ＨＣＶＲＮＡ阳性率（５０．０％）为高，其在ＰＣＲ试验中呈阳性与强阳性反应者的比率，前者亦显著高于后者（８５．２％与２５．ｏ％，Ｐ＜０．０１）。     

Hepatitis C virus markers in patients with long-term biochemical and histological remission of chronic hepatitis

We measured hepatitis C virus (HCV) RNA and antibodies against HCV recombinant proteins (C22/S1, E1/S2, E2/NS1, C33/NS3, C100/NS4, NS5) in serial serum samples from 22 interferon-treated patients with a long-term follow up (range: 36–44 months). Eleven of them showed persistently normal liver function tests and a significant histological amelioration or a complete resolution of chronic hepatitis (long-term responders, LTRs). In the remaining 11 patients (non-responders (NRs)) liver function tests normalized temporarily during therapy or remained unchanged. At the end of the follow up (3 years), viraemia was undetectable in six of 11 LTRs (54.6%). HCV-RNA was always detectable in the serum of NRs (p = 0.017). At admission, anti-C22/S1, anti-E1/S2, anti-E2/NS1, anti-C33/NS3, anti-C100/NS4 and anti-NS5 were detected in 95.4%, 40.9%, 77.3%, 95.4%, 72.7%, and 77.3% of the patients, respectively. Three years after suspension of therapy, anti-C100/NS4 was undetectable in five of six (83.3%) LTRs who cleared HCV-RNA and in only one with ongoing viraemia (20%). Anti-E2/NS1 was undetectable in 54.5% of LTRs and in no NRs (p = 0.067). Anti-E1/S2 was detected more frequently in LTRs than in NRs (81.8% vs 45.5%). Serum levels of anti-C22/S1, C33/NS3 and NS5 did not change during therapy and the follow up in either group of patients. The clearance of viraemia in LTRs was associated with that of anti-C100/NS4 (p = 0.017). Serum HCV-RNA and anti-C100/NS4 appear suitable tools for monitoring patients who respond to therapy. More than 40%) of LTRs remained HCV-RNA-positive in spite of the biochemical remission of their liver diseases.     

肝组织内丙型肝炎病毒不同抗原的定位及比较

目的研究ＨＣＶ不同抗原的组织内定位以及与病变和血ＨＣＶＲＮＡ的关系。方法５１份慢性丙型肝炎肝穿刺组织,免疫组化检测ＨＣＶＣＰ１０、ＮＳ３和ＮＳ４,其中３２份还检测ＮＳ５,同时观察组织学病变和血ＨＣＶＲＮＡ（ＲＴＰＣＲ法）。结果ＨＣＡｇ总阳性率＞６０％。不同抗原阳性率不一,以ＣＰ１０和ＮＳ５为高（各＞５０％）。阳性细胞数一般较少,散在分布;灶性集聚时,多与碎屑坏死及炎症反应相伴。几种抗原全阳性者组织病变活动度较高。血ＨＣＶＲＮＡ阳性率８０．３９％,但血与组织检测结果的重合性仅占５０．９８％;血阴性者,８／１０份组织内检出１或多种抗原。结论组织内同时检测多种抗原能提高ＨＣＡｇ检出率。     

Intrafamilial transmission of hepatitis C virus in Japan

To study the intrafamilial transmission of hepatitis C virus (HCV), 36 family members of 16 patients with anti-HCV (anti-C100-3)-positive chronic liver disease were screened for anti-HCV by an enzymelinked immunosorbent assay (ELISA). Clusters of anti-HCV -positive individuals were observed in 2 of 16 families (12.5%). Four of 35 family members (11.4%) with no history of blood transfusion were positive for anti-HCV. Two of 17 offspring (11.8%) of anti-HCV-positive females were positive for anti-HCV, while 1 of 5 spouses (20.0%) was positive for anti-HCV. These data suggest that intrafamilial transmission is one of the possible routes of infection for HCV.     

Evaluation of three hepatitis C virus-related antibodies C100, KCL-163, JCC

To evaluate the most effective method for detecting hepatitis C virus (HCV) carriers in a large population of blood donors, HCV-related antibodies were measured in 919 donor serum samples using three different enzyme-linked immunosorbent assays. The antibodies were C100 and KCL-163, nonstructural proteins of HCV, as well as JCC, a translation product of the presumptive HCV core gene. Fourteen (1.5%), 12 (1.3%), and 13 (1.4%) specimens were positive for anti-C100, anti-KCL-163, and anti-JCC, respectively. HCV RNA was detected by the polymerase chain reaction in seven (25.0%) of the 28 specimens that were anti-HCV-positive by at least one of the three assays. Four of the seven specimens were detected by anti-C100 screening, while the remaining three were not. All seven specimens were positive for KCL-163 and/or JCC antibodies. These findings suggest that screening for both KCL-163 and JCC antibodies may be of particular use in accurately identifying HCV-positive blood.This work was supported in part by a grant from the Sagawa Science Foundation.     

地高辛标记探针原位杂交检测肝组织中HCV RNA

用地高辛标记ＨＣＶ５′－ＮＣ区ｃＤＮＡ探针原位杂交，检测了２４例ＨＣＶ感染的慢性肝病患者肝组织中ＨＣＶ ＲＮＡ。结果：１７例同时检测血清抗－ＨＣＶ和ＨＣＶ ＲＮＡ均阳性患者中，１４例（８２．３％）肝组织检出ＨＣＶ ＲＮＡ。７例仅有抗－ＨＣＶ阳性患者，肝组织中未检出ＨＣＶ ＲＮＡ。ＨＣＶ ＲＮＡ特异性信号主要位于肝细胞浆。感染ＨＣＶ肝细胞周围见较多的淋巴细胞和单核细胞浸润。结果提示ＨＣＶ在肝细胞浆复