共查询到20条相似文献,搜索用时 0 毫秒
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Iushchuk ND Klimofa EA Znoĭko OO Pyleva OG Kelli EI Cheshik DS Braginskiĭ DM Lakina EI Masalova OV Kushch AA 《Klinicheskaia meditsina》2001,79(12):24-28
Puncture biopsy of the liver and blood count were made in 72 patients with chronic hepatitis C (CHC). Morphological alterations in the liver were assessed by Knodell index. The blood serum, lymphocytes and hepatic tissue were examined for a genome form of hepatitis C virus (HCV) RNA, blood lymphocytes and hepatic tissue--for a relevant replication form. HCV RNA was detected using "nested" RT-PCR. Only 26% patients had symptoms of asthenovegetative and dyspeptic syndromes. Normal alaninaminotransferase (ALT) level was observed in 24% patients, the rest had it high. HCV RNA was encounted more frequently in hepatic tissue than lymphocytes or serum (83, 68 and 46%, respectively). A replication form of HCV RNA was present in hepatic tissue of 31% patients and was absent in the lymphocytes. The incidence of the RNA detection was not related either to the disease symptoms or morphological alterations in hepatic tissue. The occurrence of the genome and replication forms in hepatic tissue does not correlate to ALT level. HCV RNA occurs more often in the serum, blood lymphocytes and in three substrates simultaneously in patients with hyperalatemia. 相似文献
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Arase Y Ikeda K Suzuki F Suzuki Y Saitoh S Kobayashi M Akuta N Someya T Hosaka T Sezaki H Kobayashi M Kumada H 《Intervirology》2005,48(6):400-404
OBJECTIVE: The aim of this study is to evaluate the serum level of KL-6 in hepatitis C virus (HCV)-positive patients with chronic liver disease. METHODS AND RESULTS: Subjects consisted of 502 HCV-positive patients. The serum samples of these patients stored at -80 degrees C were measured by enzyme-linked immunosorbent assay for KL-6 at the same time. The cutoff point of the serum KL-6 level was defined as 500 U/ml. The serum KL-6 level of the 502 patients ranged between 71 and 2,295 (median, 223) U/ml. Thirty-two of the 502 (6.4%) patients showed an elevated KL-6 level of >500 U/ml. Three of the 32 (9.4%) patients with elevated KL-6 level >500 U/ml had idiopathic pulmonary fibrosis. Multivariate analysis showed that patients achieved elevated KL-6 when: (1) they had hepatocellular carcinoma (HCC; p = 0.0007), and (2) age was >60 years (p = 0.0085). The HCC rate was 37.5% (12/32) in the patients with elevated KL-6 and 8.3% (39/470) in the patients with normal KL-6 group. The median (range) age was 70 (56-77) years in the patients with elevated KL-6 group and 60 (12-92) years in the patients with normal KL-6. CONCLUSION: The patients with HCC aged >60 years had significantly elevated serum levels of KL-6. 相似文献
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BackgroundGold nanoparticles (AuNPs) exhibit a unique phenomenon known as Surface Plasmon Resonance, which is responsible for their intense red color. This color changes to blue upon aggregation of AuNPs.ObjectiveThis work aims to develop a rapid, simple and cheap assay for direct detection of unamplified HCV RNA extracted from clinical samples using unmodified AuNPs.MethodsSerum samples were collected from healthy volunteers (n = 45) and chronic HCV patients (n = 30). Extracted RNA, hybridization buffer containing PBS, and a primer targeting the 5′UTR of HCV were mixed. The mixture was denatured, annealed, and then cooled to room temperature for 10 min followed by addition of AuNPs.ResultsSalt, primer, AuNPs concentrations and annealing temperature and time were all optimized. In HCV positive specimens, the color of the solution changed from red to blue within 1 min. The assay has a sensitivity of 92%, a specificity of 88.9%, and a detection limit of 50 copies/reaction.ConclusionsTo our knowledge, this is the first assay that allows the detection of unamplified HCV RNA in clinical specimens using unmodified AuNPs. The developed assay is highly sensitive, has a turnaround time of 30 min, and eliminates the need for thermal cycling and detection instruments. 相似文献
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目的比较两种不同核酸提取方法检测血清丙型肝炎病毒核糖核酸(HcuRNA)对临床诊断的应用价值。方法对128份HcV抗体阳性的患者血清,同时用两种实时荧光定量聚合酶链反应(PCR)方法检测HCV—RNA并检测丙氨酸氨基转移酶(ALT)水平,这两种方法分别是用二氧化硅微粒法提取RNA和纯化柱法提取RNA。结果128份HCV抗体阳性的血清中二氧化硅微粒法HCV—RNA阳性率41.41%,纯化柱法HcV—RNA阳性率59.38%,纯化枉法优于二氧化硅微粒法,差异有统计学意义(x2=8.27,P〈0.05)。49份HCV抗体阳性的血清中ALT异常,且ALT浓度变化与纯化柱法HCV—RNA水平呈正相关性(r=0.95,P〈0.05)。结论血清HCV—RNA检测对丙型肝炎诊断和病情监测均有重要的临床意义,采用纯化柱法提取RNA具有更优的临床价值。 相似文献
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Potapova AA Redchenko EB Toguzov RT Koval'chuk LV Safronova NIu Evsegneeva ZhV Shcherbo DS 《Klinicheskaia laboratornaia diagnostika》2008,(6):41-44
The purpose of the study was to elucidate whether hepatitis C virus (HCV) RNA was present in the blood of patients from a Moscow therapeutic-and-prophylactic institution with uncertain results of a study of anti-HCV that were obtained using the algorithm developed for mass screening. The two-stage scheme of testing the serum for anti-HCV, which was obligatory in accordance with the Order of the Ministry of Health of the Russian Federation, was supplemented by a study the samples having a low optical density samples and those with controversial results in the test systems with the expanded spectrum of detectable anti-HCV, the instructions of which comprises criteria for an uncertain result. Sera with uncertain results of anti-HCV tests were assessed by two polymerase chain reaction (PCR) techniques for HCV RNA. The cDNA fragments complementary to HCV RNA were detected in the sera obtained from two elderly persons in PCR when a signal was recorded in agarose gel. The other real-time PCR failed to detect RNA in none sera with an uncertain result as to anti-HCV. 相似文献
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目的 探讨HCV感染者/慢性丙型肝炎患者血清HCV-RNA含量、外周血淋巴细胞(PBL)Fas 抗原及ALT浓度的相关性.方法 对本站献血群中60例HCV感染者,50例正常者及来自本市一院56例慢性丙型肝炎患者,采用荧光定量PCR检测其HCV-RNA含量,用FACSCalibur流式细胞仪为检测外周血淋巴细胞Fas抗原,用生化自动分析仪检测ALT浓度.结果 本研究116例HCV感染者/慢性丙型肝炎患者血清HCV-RNA含量与PBL Fas抗原表达率、ALT异常率呈正相关(γ值分别为0.94、0.96,均P<0.01),但与ALT浓度无相关性γ=0.38,0.03,无统计学意义(P>0.05),ALT浓度差异无统计学意义(均P>0.05).ALT异常率差异有统计学意义(P<0.05).结论 HCV感染者/慢性丙型肝炎患者血清HCV-RNA含量与外周血淋巴细胞Fas抗原呈正相关、与ALT异常率相关性显著,外周血淋巴细胞Fas抗原与ALT异常率相关性显著. 相似文献
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两种方法检测HCV-RNA的结果分析 总被引:1,自引:0,他引:1
目的比较两种方法检测丙型肝炎病毒核酸(HCV-RNA)的结果。方法逆转录套式定性PCR(RT-nestedPCR)和荧光定量PCR对288例抗-HCV阳性的丙型肝炎患者血清标本进行HCV-RNA检测。结果288例抗-HCV阳性血清标本有248例RT-nested-PCR结果阳性,阳性率为86.1%(248/288);226例荧光定量PCR检测HCV-RNA>103拷贝/ml,检出率为78.5%(226/288),两种检测结果的阳性符合率为85.9%(219/255)。同时,29例标本定性结果阳性,定量结果<103拷贝/ml;7例标本定性结果阴性,定量结果>103拷贝/ml,差异具有统计学意义(χ2=12.25,P<0.01)。结论两种方法检测HCV-RNA,可以提高检测的灵敏度,为临床治疗提供依据。 相似文献
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HCV感染者/慢性丙型肝炎患者血清HCVRNA含量、外周血淋巴细胞Fas抗原及ALT浓度的相关性 总被引:1,自引:0,他引:1
目的探讨HCV感染者/慢性丙型肝炎患者血清HCV-RNA含量、外周血淋巴细胞(PBL)Fas抗原及ALT浓度的相关性。方法对本站献血群中60例HCV感染者,50例正常者及来自本市一院56例慢性丙型肝炎患者,采用荧光定量PCR检测其HCV-RNA含量,用FACSCalibur流式细胞仪为检测外周血淋巴细胞Fas抗原,用生化自动分析仪检测ALT浓度。结果本研究116例HCV感染者/慢性丙型肝炎患者血清HCV-RNA含量与PBLFas抗原表达率、ALT异常率呈正相关(γ值分别为0.94、0.96,均P﹤0.01),但与ALT浓度无相关性γ=0.38,0.03,无统计学意义(P均>0.05),ALT浓度差异无统计学意义(均P>0.05),ALT异常率差异有统计学意义(P﹤0.05)。结论HCV感染者/慢性丙型肝炎患者血清HCV-RNA含量与外周血淋巴细胞Fas抗原呈正相关、与ALT异常率相关性显著,外周血淋巴细胞Fas抗原与ALT异常率相关性显著。 相似文献
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Direct detection of hepatitis C virus (HCV) RNA in serum or plasma is useful for validating the performance of anti-HCV assays and for the discrimination of persons with persistent HCV infections from those with resolved infections. Quantitation of HCV RNA may also be useful for disease prognosis and therapeutic monitoring. Previous studies have reported detection of HCV RNA in 50 to 70 percent of blood donors who were positive on anti-HCV supplemental tests. There is concern that specimen processing and storage conditions might influence the stability, and hence the detectability, of HCV RNA. To address this concern, the rate of detection of HCV RNA by the polymerase chain reaction (PCR) using donor pilot tube sera (PTS) previously subjected to routine donor screening and supplemental testing was compared with HCV PCR results obtained with fresh-frozen plasma (FFP) derived from the same donations. All 16 anti-HCV supplemental test-positive donations evaluated were HCV RNA positive with FFP, whereas only 10 (62.5%) were positive with PTS (p = 0.024). None of 11 FFP or PTS samples from HCV enzyme immunoassay-reactive donations not confirmed by supplemental anti-HCV assays tested positive for HCV RNA. Direct comparison of sample type (serum vs. plasma) and various storage conditions using specimens from two seropositive donors showed that room-temperature storage results in marked reduction in HCV RNA signal, while replicate freezing and thawing caused a moderate reduction. These data indicate that well-controlled sample processing and storage conditions are critical to the sensitive and potentially quantitative analysis of HCV RNA. 相似文献
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目的探讨丙型肝炎病毒核心抗原(HCV-cAg)对丙型肝炎(简称丙肝)筛查的意义。方法收集2014年10月至2015年10月8 000例门诊及住院患者,用酶联免疫吸附试验(ELISA)检测HCV-cAg、丙型肝炎病毒抗体(HCV-Ab),并对HCVcAg或HCV-Ab阳性标本采用聚合酶链反应(PCR)法检测HCV-RNA进行确诊。结果以HCV-cAg或HCV-Ab阳性为标准,从8 000例血清样本中初步筛查出阳性样本82例,经HCV-RNA确证,其中HCV-RNA阳性73例,阴性9例,HCV-cAg的灵敏度及特异度分别为45.83%和99.98%,HCV-Ab的灵敏度及特异度分别为94.44%和99.90%,联合检测HCV-cAg与HCV-Ab的灵敏度及特异度分别为100.00%和99.86%。结论在丙肝筛检工作中,HCV-cAg与HCV-Ab二者有互补性,联合检测HCV-cAg与HCV-Ab有助于提高HCV筛查率。 相似文献
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2000~2002年丙型肝炎病毒核酸逆转录聚合酶链反应测定室间质量评价 总被引:2,自引:1,他引:2
目的 评价2000年至2002年临床基因扩增检验实验室在丙型肝炎病毒(HCV)RNA逆转录-聚合酶链反应(RT-PCR)检测中的质量。方法 从2002年至2003年每年进行质评两次,每次发放HCVRNA样本5份,其中阳性样本3—4份,含量在10^4-10^7拷贝数/ml范围内。阴性样本1~2份,均为冻干品。要求各个实验室按规定时间检测并回报结果,然后统计分析。结果 对质评样本测定全部正确的比率从2000年的001批的53.3%(24/45)、002批的4.8%(2/42)和2001年011批的73.8%(31/42)、012批的79.2%(42/53),上升到2002年021批的89.7%(105/117)和022批的91.6%(109/119)。假阳性率也从2000年和2001年的高至26.7%和14.3%降至2002年的8.0%以下。对在10^5和10^4拷贝数/ml数量级的质评样本,测定假阴性率从2000年的15.6%(0011)、19.0%(0025)和81.0%(0023)下降到2001年的11.9%(0112)、11.3%(0125)和9.5%(0115),到2002年更是下降到了4.6%(0221)、3.7%(0223)和3.8%(0212)。从试剂盒的临床使用评价来看,从2000年到2002年各厂家试剂的临床检测特异性、灵敏度和符合率均有明显改善。结论 从2000-2002年全国临床HCV RNA RT-PCR测定假阳性和假阴性明显降低。说明所建立的HCV RNA临床RT-PCR测定室间质量评价项目能很好的监测实验室存在质量问题,从而促使其加以改进。 相似文献
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Koppelman MH Assal A Chudy M Torres P de Villaescusa RG Reesink HW Lelie PN Cuypers HT 《Transfusion》2005,45(8):1258-1266
BACKGROUND: The performance of the recently launched Procleix Ultrio (Chiron/Gen-Probe) human immunodeficiency virus-1 (HIV-1), hepatitis C virus (HCV), and hepatitis B virus (HBV) blood screening assay was evaluated in a European multicenter study. STUDY DESIGN AND METHODS: Serial dilutions of reference materials were tested to determine the detection limits. Robustness and specificity were assessed by testing alternating high-load HCV RNA-positive and -negative samples, and 2912 test pools of eight donations. The added value of minipool and single-donation HBV nucleic acid testing protocols was compared to the currently used Prism (Abbott GmbH & Co. KG) hepatitis B surface antigen (HBsAg) and Auszyme (Abbott GmbH & Co. KG) dynamic HBsAg tests in 15 HBV seroconversion panels. RESULTS: The 95 percent detection limits (and 95% confidence interval [CI]) on the WHO International Standards was 26 (16-58) IU per mL for HIV-1 RNA, 4.6 (3.7-6.5) IU per mL for HCV RNA, and 11 (7.3-22) IU per mL for HBV DNA. No cross-contamination was observed. Testing 2912 pools of eight donations revealed 16 initial reactive samples; 11 were confirmed. The specificity after initial testing and percentage of invalid results were 99.83 and 0.48 percent, respectively. The HBV window-period (WP) reductions relative to HBsAg seroconversion in Prism and Auszyme dynamic HBsAg were, respectively, 6 days (95% CI, 3-8) and 9 days (95% CI, 7-12) in 1:8 minipool (MP) testing. CONCLUSION: The performance characteristics of Procleix Ultrio assay and the Procleix HIV-1 and HCV assay are comparable. The sensitivity for HIV-1 and HCV met the directives of the Paul-Ehrlich Institute and the FDA. The assay can reduce the WP for HBV by 6 days to 2 weeks when used in small MP (<1:8) or single-donation screening protocols. 相似文献
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BACKGROUND: Recently developed nucleic acid testing (NAT) assays incorporating simultaneous detection of human immunodeficiency virus (HIV), hepatitis C virus (HCV), and hepatitis B virus (HBV) have made HBV NAT screening more feasible for blood services. This study compared the performance of two "multiplex" NAT assays and their automated testing platforms. STUDY DESIGN AND METHODS: The HBV NAT yield rate was estimated by testing 10,397 Hong Kong (HK) donor samples concurrently on the PROCLEIX ULTRIO (Ultrio) assay as individual donor samples with the TIGRIS and on the cobas TaqScreen multiplex (cobas MPX) test in pools of 6 with the cobas s 201. Analytical sensitivity was assessed by probit analysis of diluted international standards and operational performance was compared. RESULTS: Each system detected two different HBV NAT yield samples for a combined rate of 0.04 percent. One additional sample was reactive on the cobas MPX test but remained unresolved. The 95 percent detection limits for HIV-1, HBV, and HCV were 42.2, 12.2, and 2.0 IU per mL, respectively, for Ultrio and 50.5, 8.4, and 6.0 IU per mL for the cobas MPX. The invalid test and failed run rates were 0.05 and 2.92 percent, respectively, for the TIGRIS and 2.39 and 5.53 percent for the cobas s 201. CONCLUSION: Clinical sensitivity for HBV in HK blood donors was equivalent, as was the analytical sensitivity for HIV-1 and HBV; however, the Ultrio assay had a higher analytical sensitivity for HCV. Despite a shorter downtime and mean time of repair for the cobas s 201, the TIGRIS demonstrated better overall operational performance. 相似文献
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Recombinant immunoblot assay reaction patterns and hepatitis C virus RNA in blood donors and non-A, non-B hepatitis patients 总被引:5,自引:0,他引:5
D Bresters ; HL Zaaijer ; HT Cuypers ; HW Reesink ; IN Winkel ; PJ van Exel-Oehlers; AA van Drimmelen; PL Jansen ; CL van der Poel; PN Lelie 《Transfusion》1993,33(8):634-638
To establish the value of the second-generation recombinant immunoblot assay (RIBA-2) and cDNA polymerase chain reaction (cDNA PCR) for confirmation of hepatitis C virus (HCV) infection, anti-HCV reaction patterns and the presence of HCV RNA were examined in 610 blood donors and 255 non-A, non-B hepatitis patients who were positive or indeterminate in RIBA-2. Of RIBA-2-positive donors (n = 191) and patients (n = 224), 75.4 and 89.7 percent, respectively, were HCV RNA positive. The most frequently observed anti-HCV recognition patterns in HCV RNA-positive donors and patients were c22, c33c, and c100 and/or 5- 1-1 (67.3%, 57.7%) and c22, c33c (24.8%, 29.3%). Among subjects with a RIBA-2-indeterminate result, HCV RNA was detected more often in patients (n = 31) than in donors (n = 419): 67.7 and 2.1 percent, respectively. In viremic persons with single-band reactivity in RIBA-2, this reactivity was always directed against either c22 or c33c. HCV RNA was detected by cDNA PCR in none of 162 persons with only anti-c100 and/or anti-5-1-1 reactivity. Therefore, RIBA-2 reactivity against c100 in combination with 5-1-1 should not be considered positive but indeterminate. In RIBA-2-indeterminate persons, HCV RNA detection is necessary for reliable confirmation of HCV infection. 相似文献