NAT results of HBV and HCV under different vacuum collection tubes, storage temperature and storage time / 中国输血杂志

【Objective】 To verify the results of HBV DNA and HCV RNA screening under different brands of vacuum collection tubes for blood samples, storage temperature and storage time. 【Methods】 Experiment 1 was conducted as follows: blood samples were collected simultaneously from 52 voluntary blood donors using two brands(divided into group A and group B) of vacuum collection tubes for blood samples. The plasma separation of group A and group B were compared, and the effects of storage time on the NAT yield of HBV DNA and HCV RNA were statistically analyzed. Experiment 2 was conducted as follows: the effects of different storage temperature, time and tubes on the NAT yield of HBV DNA and HCV RNA samples with low viral load in group A and B were verified and compared in the simulated phlebotomy condition. 【Results】 In Experiment 1: After centrifugation, blood plasma layer and cells layer were separated completely in group A(100%, 52/52), but one sample was not well separated in group B(1/52, 1.92%). After 4 to 10 h after collection, blood samples of two groups were centrifuged and screened for HBV DNA, HCV RNA within 24 h. No positive samples were yielded and the Ct values of internal control(IC-DNA and IC-RNA) were uniform. In Experiment 2: Whole blood samples, stored for either 4 h or 6~10 h at 4 ℃ or 25℃ before centrifugation, showed no difference on the NAT-yield of HBV DNA nor HCV RNA samples with low viral load(P>0.05). Ct values of HBV DNA and HCV RNA of group A was similar to those of group B as centrifuged samples were stored for 24 h or 72~104 h at 4℃(P>0.05), but all increased as the storage time prolonged. Ct values of HBV DNA in group A increased from 33.45±0.29(24 h) to 33.82±0.08(72~104 h) and HCV RNA from 35.21±0.20 to 36.12±0.43; HBV DNA from 33.46±0.25 to 34.30±0.60 and HCV RNA from 35.47±0.24 to 36.49±0.51 in group B. 【Conclusion】 Under certain laboratory condition, different storage time, storage temperature and tubes shed few effect on the NAT-yield of HBV DNA and HCV RNA samples with low virus loads. However, it is suggested that the blood sample be detected within 72 h after centrifugation at 4 ℃ storage.

Epidemiological characteristics of occult hepatitis C virus and safety of blood transfusion / 中国输血杂志

Occult hepatitis C infection (OCI) is defined as HCV RNA not detected in serum or plasma but in hepatocytes and peripheral blood mononuclear cells (PBMCs). OCI exists in general population and voluntary blood donors, and its infectivity and risk of transmission by transfusion has been confirmed. HCV RNA in PBMCs could not be detected in plasma or serum by blood screening in transfusion services, neither by enzyme-linked immunosorbent assay nor by nucleic acid amplification testing. OCI has become a potential threat to transfusion safety, therefore effective detection technologies and transmission blocking strategies need to be further developed.

Exploration of an Efficient Simultaneous Molecular Detection Method of HIV, HCV, and Syphilis from a Single Dried Blood Spot / 生物医学与环境科学（英文）

Objective@#The aim of the present study was to evaluate the performance of the simultaneous detection of HIV-1 RNA, HIV-1 DNA, and HCV RNA using one dried blood spot (DBS) as an alternative sample to plasma.@*Method@#A total of 571 paired DBS/plasma samples were collected from men who have sex with men (MSM) and injection drug users (IDUs), and serological and molecular assays were performed. Using plasma results as the reference standard, the performance of DBS tests for HIV-1 RNA, HIV-1 DNA, and HCV RNA was evaluated. Pearson's correlation coefficients and Bland-Altman analysis were performed to assess the correlation and concordance between DBS and plasma.@*Results@#Among paired plasma/DBS samples with detectable HIV-1 RNA and HCV RNA, five samples (5/32) were not detectable in DBS, while measurable HIV-1 RNA levels were present in plasma (1.44 to 3.99 log @*Conclusion@#The performance of the simultaneous detection of HIV-1 RNA, HIV-1 DNA, and HCV RNA using one DBS was acceptable. DBS, as an alternative sample to plasma, may be a viable option for the simultaneous detection of HIV-1 RNA, HIV-1 DNA, and HCV RNA in resource-limited settings or for individuals living in areas that are difficult to access.

Analysis of hepatitis C virus 5′ non-coding region for viral genotyping and the level of viral replication / 中华实验和临床病毒学杂志

Objective@#To investigate hepatitis C virus(HCV)genotyping and the serum HCV-RNA concentration in patients infected with different HCV genotypes and to provide information for evaluation of disease condition and anti-viral treatment efficacy.@*Methods@#A total of 60 anti-HCV positive serum samples were collected before antiviral treatment. RT-PCR was performed for the 5′ non-cording region and was followed by nucleotide sequencing for HCV genotyping. Meanwhile, serum HCV-RNA concentration was detected by quantitative PCR. SPSS21.0 and Graphpad Prism 5.0 software were used for data analysis. Analysis of variance (ANOVA) was used for comparison among multi-groups and the t-test was used for comparison between two groups.@*Results@#The frequencies of HCV genotypes 1b, 3a, 1a and 2a were 48.3% (29/60), 23.3% (14/60), 16.7% (10/60) and 10% (6/60), respectively. And, there is one subtype 2c was detected in this study. The mean serum viral concentration with standard deviation of HCV in genotype 1a, 1b, 2a, and 3 a were 5.46±1.19, 6.22±0.78, 5.47±0.65, and 5.38±0.98 log10 (IU/ml) respectively.@*Conclusions@#The infection rate of HCV genotype 1 was significantly higher than that of genotype 2 and 3 (P<0.01). The statistical analysis showed the serum HCV-RNA concentration in patients with subtype 1b was significantly higher than that of the subtype group 1 a, 2 a, and 3 a (P<0.05). The study of the relationship between HCV genotypes and the serum HCV-RNA concentration may contribute to anti-viral treatment prescription for hepatitis C patients.

Comparison of TRIzol method andmagnetic beads method in quantitative detection of hepatitis C virus RNA / 第二军医大学学报

Objective To compare the effects of TRIzol and magnetic beadsmethods on quantitative detection of hepatitis C virus (HCV) RNA. Methods Serum samples and genotype information of 117 patients with positive HCV infection were collected. HCV RNA was extracted from serum samples by TRIzol method and magnetic bead method, respectively. And then the viral load of HCV RNA was detected by quantitative PCR to compare the difference between the two methods. Results qPCRresults showed that a good linear correlation existed between TRIzol and magnetic beads methods: y=0. 978x+0. 063 (R2=0. 973). Bland-Altman statistical analysis showed that the average logarithmic value of viral load ofHCV RNA extracted by TRIzol method was slightly lower than that of magnetic beads method, without significant difference (P>0. 05). There were no significant difference among the genotypes 1a, 1b, 2a, 3a or 6a between the two methods (P>0. 05). Conclusion TRIzol method is comparable to magnetic beads method in HCV RNA quantitative detection, with less samplevolume and lower cost, indicating that t might be widely used for developing ktt and HCV RNA clinical detection in China.

The value of Anti-HCV-IgG antibody and serum HCV-RNA in the diagnosis of hepatitis c / 国际检验医学杂志

Objective To study the anti-HCV-IgG antibody and serum HCV-RNA value in the diagnosis of hepatitis c.Methods From December 2015 to December 2006,76 patients with hepatitis C were enrolled in this study.All patients were tested for serum anti-HCV-IgG antibody and serum HCV-RNA,and the positive detection rate of hepatitis C was detected by two methods alone.Results 76 specimens of anti-HCV IgG antibody testing positive rate was 27.63％,the detection of serum HCV-RNA positive rate was 25.00％,the two kinds of joint detection positive rate was 22.37％,the three methods positive rate were relatively close,but there was no significant difference(P＞0.05).According to different loads interval differentiate HCV-RNA virus detection rate of rising,anti-HCV-IgG five interval were 60.00 ％,83.33 ％,100.00 ％,100.00 ％,100.00 ％.Conclusion The detection of single anti-HCV IgG antibody or serum HCV RNA in the diagnosis of hepatitis c have limitations,the joint detection can effectively reduce the missed diagnosis and improve positive rate and provide a reliable basis for clinical diagnosis of hepatitis C.

The value of Anti-HCV-IgG antibody and serum HCV-RNA in the diagnosis of hepatitis c / 国际检验医学杂志

Objective To study the anti-HCV-IgG antibody and serum HCV-RNA value in the diagnosis of hepatitis c.Methods From December 2015 to December 2006,76 patients with hepatitis C were enrolled in this study.All patients were tested for serum anti-HCV-IgG antibody and serum HCV-RNA,and the positive detection rate of hepatitis C was detected by two methods alone.Results 76 specimens of anti-HCV IgG antibody testing positive rate was 27.63％,the detection of serum HCV-RNA positive rate was 25.00％,the two kinds of joint detection positive rate was 22.37％,the three methods positive rate were relatively close,but there was no significant difference(P＞0.05).According to different loads interval differentiate HCV-RNA virus detection rate of rising,anti-HCV-IgG five interval were 60.00 ％,83.33 ％,100.00 ％,100.00 ％,100.00 ％.Conclusion The detection of single anti-HCV IgG antibody or serum HCV RNA in the diagnosis of hepatitis c have limitations,the joint detection can effectively reduce the missed diagnosis and improve positive rate and provide a reliable basis for clinical diagnosis of hepatitis C.

Development of a New TaqMan-MGB Probe-Based Real-Time qPCR Method for the Detection of HCV-RNA / 现代检验医学杂志

Objective Developing a rapid and accurate real-time qPCR method for the detection of HCV-RNA.Methods HCV nucleotide sequence was analysed in Clustal software and primers and probe were designed in the conserved region of 5'UTR.The reaction system optimization of real-time qPCR method was used chessboard titration,pseudoviral particles were used as quantitative standard to assess the performance.New methods was compared with clinical commonly used kit of HCV-RNA and discuss the application value.Results The sensitivity of new real-time qPCR method was 50 IU/ml,coefficient variation was less than 5％.The quantitative results of this method could be traceable to national standards of GBW09151a.40 samples were determined by new methods and clinical commonly used kit of HCV-RNA,the positive concordance rate was 100 ％,the negative concordance rate was 56 ％.14 samples were positive by new method,but negative by Qiagen kit,illustrating that the sensitivity of new method was superior to Qiagen kit.Conclusion New TaqMan-MGB probe-based real-time qPCR method is a specific,sensitive,simple,rapid and exactly used to detection of HCV-RNA.

Relevance of Serum HCV-RNA Level with Routine Liver Function Indices and Blood Routine Parameters in Patients with Hepatitis C / 现代检验医学杂志

Objective To explore the correlation of serum HCV-RNA level with liver function indices and blood routine parameters in patients with hepatitis C,and provide the references for the early diagnosis and monitoring the prognosis of hepatitis C.Methods The study population comprised 651 patients who were diagnosed with hepatitis C in the Second Affiliated Hospital of Medical College of Xi' an Jiaotong University.Serum level of HCV RNA was determined by quantitative realtime polymerase chain reaction (real-time PCR).The data obtained were divided into six groups according to the result of HCV RNA level.Then the correlation of serum HCV-RNA level with liver function indices and blood routine parameters was analyzed.Results With the rising level of serum HCV-RNA,the levels of liver function indices including ALT,AST,AST/ALT,GGT and TBIL were all significant increased in hepatitis C patients.However,A/G presented a declining trend with the increase of serum HCV-RNA level.The lowest value and highest value of ALT,AST,AST/ALT,GGT,TBIL and A/G were 32.3±9.7 U/L and 96.2±13.6 U/L,31.1±8.38 U/L and 113.5±15.9 U/L,0.86±0.09 and 1.19±0.11,29.2± 14.5 U/L and 52.7±16.2 U/L,17.2±4.32 μmol/L and 26.0±5.58 μmol/L,0.98±0.07 and 1.35±0.14,respectively.Significant differences in ALT,AST,AST/ALT,GGT,TBIL and A/G were observed among these groups (F value was 457.1,656.4,149.1,40.18,46.56 and 146.98,respectively.all P＜0.01).Moreover,the levels of blood routine parameters including WBC,NEUT％,RBC,H b and PLT were all significant decreased while that of LY％ was increased with the rising level of serum HCV-RNA in hepatitis C patients.The lowest value and highest value of WBC,NEUT％,LY％,RBC,Hb and PLT were (4.30±0.22) ×109/L and (6.02±0.27) × 109/L,(48.13±3.56)％ and (59.28±3.40)％,(31.05±3.41) ％ and (38.81 ± 4.65)％,(3.73 ± 1.70) × 1012/L and (4.65± 1.88) × 1012/L,(122.01±5.58) g/L and (135.37 ±8.50)g/L,(102.65± 16.87) × 109/L and (148.21 ± 14.40) × 109/L,respectively.There was no significant difference in RBC among groups (F=1.926,all P＜0.05).However,the differences in other blood routine parameters than RBC among groups were statistically significant (F value of WBC,NEUT％,LY％,Hb and PLT was 349.0,132.145,43.65,38.91 and 52.21,respectively.all P＜0.01).Conclusion Serum HCV-RNA level was correlated with routine liver function indices and blood routine parameters.It is important to perform the combined detection of serum HCV-RNA level,routine liver function indices and blood routine parameters in hepatitis C patients,which can help to guide diagnosis and to evaluate the prognosis.

Clinical Application Value of the Result of the HCV RNA,FIB-4 and Levels of 2 5-Hydroxy Vitamin D for Patients with Hepatitis C / 现代检验医学杂志

Objective To analyze the effects of 25-hydroxyvitamin D[25(OH)D]on the result of the HCV RNA and the FIB-4 in the patients with hepatitis C.Methods 255 serum samples were random collected from the patients with hepatitis C and 218 serum samples were random collected from the healthy people.The 25(OH)D,HCV RNA,aspartate aminotransferase (AST),alanine aminotransferase (ALT)and blood platelet (PLT)were detected.Then,compared the results of the 25 (OH)D in the patients with hepatitis C and the healthy group.Analyzed the relevance between the concentration of 25(OH) D and HCV RNA.According to the quartile concentration of the 25(OH)D,the patients with hepatitis C were categorized to four groups.The relationship of FIB-4 between HCVRNA and 25(OH)D was analyzed.Results The average concentration of the 25(OH)D in the patients with hepatitis C and healthy people were 48.16±1.41 nmol/L vs 60.42±1.34 nmol/L, with a significant difference (t=4.682,P<0.01).There were 38 patients (14.90%)had severe deficiency of 25(OH)D (<25 nmol/L)in 255 patients with hepatitis C.And there were 8 patients (3.67%)had severe deficiency of 25(OH)D (<25 nmol/L)in 218 healthy people,with a significant difference (t=5.216,P<0.01).Then found no relevance between the log-arithmic of the HCV RNA and the concentration of the 25(OH)D (r2=0.018 8,P=0.412)and there was significant differ-ence between the proportion of FIB-4 in the highest quartile concentration of the 25(OH)D and the lowest quartile concen-tration of the 25(OH)D (χ2=8.190,P=0.042).Conclusion The patients with hepatitis C were easier to have a severe de-ficiency of 25(OH)D than the healthy people.The hepatitis C patients should been suggested to supply the vitamin D.FIB-4 has a significant difference with 25(OH)D and no great effects on the result of the HCV RNA.

Level diversifies of plasma CCL19 and CCL21 in male drug users infected HCV / 中国免疫学杂志

Objective:To analyze the level diversifies of plasma CCL19 and CCL21 in the male drug users infected HCV.Methods: The plasma CCL19 and CCL21,anti-HCV and HCV RNA were detected by ELISA quantitation,ELISA qualitation and Real-time RT-PCR respectively.Compared with 60 healthy man,the level diversifies of plasma CCL19 and CCL21 in 391 male drug users conducted as part of HCV Surveillance Programme in Zhuhai were analyzed.Results: 180 of 391 male drug users were infected HCV and the infection rate was 46.04%.The level of plasma CCL19 and CCL21 in the male drug users[anti-HCV(-)/HCV RNA(-)] were higher than that in the healthy man (P≤0.001).The level of plasma CCL19 and CCL21 in anti-HCV(-)/HCV RNA(+) group was lower than that in the others(P≤0.05) .Compared with that in the healthy man,the level of plasma CCL19 and CCL21 in the drug abuse anti-HCV(+)/HCV RNA(-) group had significant deviation(P<0.05).Conclusion: Drug abuse can heighten the level of plasma CCL19 and CCL21 in man.Increasing the level of plasma CCL19 and CCL21 may conduce to spontaneous HCV clearance.It may prognosticate that HCV infection will be persistent and have a bad consequence when the level of plasma CCL19 and CCL21 were degraded.

Clinical Utility of a New Automated Hepatitis C Virus Core Antigen Assay for Prediction of Treatment Response in Patients with Chronic Hepatitis C

Hepatitis C virus core antigen (HCV Ag) is a recently developed marker of hepatitis C virus (HCV) infection. We investigated the clinical utility of the new HCV Ag assay for prediction of treatment response in HCV infection. We analyzed serum from 92 patients with HCV infection who had been treated with pegylated interferon and ribavirin. HCV Ag levels were determined at baseline in all enrolled patients and at week 4 in 15 patients. Baseline HCV Ag levels showed good correlations with HCV RNA (r = 0.79, P < 0.001). Mean HCV Ag levels at baseline were significantly lower in patients with a sustained virologic response (SVR) than in those with a non SVR (relapse plus non responder) based on HCV RNA analysis (2.8 log10fmol/L vs. 3.27 log10fmol/L, P = 0.023). Monitoring of the viral kinetics by determination of HCV RNA and HCV Ag levels resulted in similarly shaped curves. Patients with undetectable HCV Ag levels at week 4 had a 92.3% probability of achieving SVR based on HCV RNA assay results. The HCV Ag assay may be used as a supplement for predicting treatment response in HCV infection, but not as an alternative to the HCV RNA assay.

Detection of hepatitis C virus RNA in the peripheral blood mononuclear cells in patients with chronic hepatitis C and its relationship with effect of anti-viral treatment / 中华传染病杂志

Objective To detect the change of hepatitis C virus (HCV)RNA in the peripheral blood mononuclear cells (PBMC)and serum of patients with chronic hepatitis C (CHC)during treatment with peg-interferon α-2a (Peg IFNα-2a)plus ribavirin (RBV),and to analyze the clinical significance of HCV RNA detection in PBMC.Methods The peripheral blood samples of 20 CHC patients who visited Department of Infectious Diseases in Guangzhou No.8 People′s Hospital from June 2013 to December 2014,were collected during treatment with Peg IFNα-2a+RBV at different time points (week 0,2,4, 12,24,36 and 48).Serum and PBMC were separated.Accurate fluorescence quantification assay (Cobas TaqMan real time polymerase chain reaction[PCR])was used to detect HCV RNA level in serum,while real-time PCR and nest-PCR were applied to detect HCV RNA in PBMC.Categorical data were analyzed byχ2 test.Results Accurate fluorescence quantification of serum HCV RNA showed that HCV RNA level decline rapidly after treatment (F = 148.06,P < 0.01 ),and 18 patients achieved HCV RNA undetectable at week 12 of treatment.The positive rate of nest-PCR was higher than real-time PCR (all P <0.01).Comparison of HCV RNA levels in serum and PBMC from 20 cases found that,the clearance rate of HCV RNA in PBMC was postponed.Two patients whose HCV RNA in PBMC kept detectable relapsed at week 24 after end of treatment.Conclusions HCV RNA can be detected in PBMC of CHC patients and the positive rate of nest-PCR is higher than real-time PCR.Antiviral therapy is effective on HCV both inside and outside PBMC,but the clearance rate of HCV RNA in PBMC is postponed compared with that in serum.Slow clearance of HCV in PBMC may be a risk factor for relapse after end of treatment.

Correlation Analysis of HCV-RNA,HCV-Ab and HCV-cAg / 现代检验医学杂志

Objective To investigate the correlation of HCV-RNA with detection indexes HCV-Ab and HCV-cAg in its clini-cal application effect among patients with hepatitis C.Methods HCV-cAg and HCV-Ab in 140 cases of HCV-RNA were detected by enzyme linked immunosorbent assay in cases of PCR,which were detected by real-time fluorescence quantitative PCR.Results 127 cases in 140 cases of HCV-RNA positive serum were HCV-cAg positive,in line with the rate of 90.71%,and the cases of 110 HCV-Ab positive,in line with the rate of 78.57%.The positive detection rate of HCV-cAg with different HCV-RNA concentration was increased with the increase of HCV virus content,and the serum of different HCV-RNA concentration had no significant changes in HCV-Ab detection results.Conclusion The detection results of HCV-cAg had a high coincidence rate with HCV-RNA.Therefore detection of HCV-cAg can be as a complementary detec-tion of HCV-Ab,as the window period of HCV infection and infection in immunocompromised persons screening provides a simple,inexpensive method.At the same time it provides rapid screening for HCV infection provide diagnostic basis for those basic medical units who do not have the conditions for detection of HCV-RNA.

Response of peg IFN and ribavarin in HIV-HCV co-infection.

Background: Hepatitis C infection is one of most common co-infection in HIV. HIV infection influences the natural evolution of chronic hepatitis by higher rate of viral persistence, accelerating fibrosis, cirrhosis progressing to end-stage liver disease. Objective: This retrospective study was conducted in order to see the response to Peg- IFN α-2b with Ribavarin in HIV HCV co-infection. Material and Methods: Alanine aminotransferase and Aspartate Aminotransferase, HCV RNA quantitative and Genotype study, Fibroscan, CD4 were collected from the medical records of ART centre. Results: The mean baseline viral load (log10) was 5.76, 3.00 at 1st month, 0.44 at 3rd month, 0 at 6th month and 12th month. RVR was observed in 77.7%, EVR in 88.8%, SVR of 100% at 6th and 12th month. The mean OT and PT reduction at 3rd month was 116.11(57.79%) and 132 (61.68%) respectively, at 6 month was 158 (78.65%) and 177.56 (82.97%) respectively, at 12th month was 159(79.14%) and 176.56 (82.50%). Fibrosis at the start of treatment was 19.0 KPa, 10.00 KPa at 6th month and 8.20 KPa at 12th month. Conclusion: Study shows that SVR can be achieved in HCV HIV co infected patients with IFN and Ribavarin therapy which in turn reduces the morbidity and mortality due to liver disease. Inspite of virological response, few patients continue to have deranged AST and ALT and progressive liver fibrosis.

Indeterminate RIBA results were associated with the absence of hepatitis C virus RNA (HCV-RNA) in blood donors

Introduction: Hepatitis C virus (HCV) infection is diagnosed by the presence of antibodies and is supplemented by confirmatory testing methods, such as recombinant immunoblot assay (RIBA) and HCV-RNA detection. This study aimed to evaluate the efficacy of RIBA testing to diagnose HCV infection in blood donors positive for anti-HCV antibodies. Methods: A total of 102 subjects positive for anti-HCV determined by enzyme-linked immunosorbent assay (ELISA) at the Hematology and Hemotherapy Foundation of Bahia (HEMOBA) were later assessed with new samples using the Abbott Architect anti-HCV test (Abbott Diagnostics, Wiesbaden, Germany), the RIBA III test (Chiron RIBA HCV 3.0 SIA, Chiron Corp., Emeryville, CA, USA), the polymerase chain reaction (PCR; COBAS® AMPLICOR HCV Roche Diagnostics Corp., Indianapolis, IN, USA) and line probe assay (LiPA - Siemens, Tarrytown, NY, USA) genotyping for HCV diagnosis. Results: Of these new samples, 38.2% (39/102) were positive, 57.8% (59/102) were negative and 3.9% (4/102) were indeterminate for anti-HCV; HCV-RNA was detected in 22.5% (23/102) of the samples. RIBA results were positive in 58.1% (25/43), negative in 9.3% (4/43) and indeterminate in 32.6% (14/43) of the samples. The prevailing genotypes were 1 (78.3%, 18/23), 3 (17.4%, 4/23) and 2 (4.3%, 1/23). All 14 samples with indeterminate RIBA results had undetectable viral loads (detection limit ≤50 IU/mL). Of these samples, 71.4% (10/14) were reevaluated six months later. Eighty percent (8/10) of these samples remained indeterminate by RIBA, and 20% (2/10) were negative. Conclusions: In this study, individuals with indeterminate RIBA results had no detectable HCV-RNA. .

To estimate the significance of anti-HCV, AST, ALT in predicting hepatitis C viremia using receiver-operating characteristic (ROC) cure and Logistic regression / 中国医师杂志

Objective To evaluate the usefulness of anti -HCV signal-to-cutoff (S/CO)ratio, AST, ALT and the combined examination of anti-HCV S/CO, AST, ALT for predicting HCV RNA results by a model of logistic regression and receiver -operating characteristic (ROC) curve.Methods Five hundred and eighty -eight anti-HCV positive samples were tested by ELISA , followed by RT-PCR to detect HCV-RNA and enzyme rate method to detect AST, ALT.Patients were divided into viremia and non -viremia groups according to HCV-RNA results.Logistic regression and ROC curve analysis was performed to evaluate the diagnostic accuracy of each index for a diagnosis of viremia.Results The serum anti-HCV S/CO ratio, AST, ALT of HCV-RNA positive group were higher than HCV-RNA negative group, showing significant statistical difference ( P AST >ALT.The Area Under Curve(AUC) of the combined examina-tion of anti-HCV S/CO ratio, AST and ALT was 0.949(95% confidence interval,0.932 to 0.966), higher than the AUC of anti-HCV S/CO ratio, AST and ALT single index examinations , which was 0.894(95 % confidence interval, 0.862 to 0.926), 0.823(95%confidence interval, 0.789 to 0.856) and 0.788(95% confidence interval, 0.750 to0.826 ) respectively.C onclusions The diag-nostic relevance of the three biochemical markers for predicting the presence of viremia was anti -HCV S/CO ratio >AST >ALT.The combined examination of anti-HCV S /CO ratio, AST and ALT in predicting hepatitis C viremia is superior to any single index examina -tion and it can increase the detecting ability of HCV -RNA greatly.

Predictors of spontaneous viral clearance and outcomes of acute hepatitis C infection

BACKGROUND/AIMS: This study evaluated the predictors of spontaneous viral clearance (SVC), as defined by two consecutive undetectable hepatitis C virus (HCV) RNA tests performed > or =12 weeks apart, and the outcomes of acute hepatitis C (AHC) demonstrating SVC or treatment-induced viral clearance. METHODS: Thirty-two patients with AHC were followed for 12-16 weeks without administering antiviral therapy. RESULTS: HCV RNA was undetectable at least once in 14 of the 32 patients. SVC occurred in 12 patients (37.5%), among whom relapse occurred in 4. SVC was exhibited in 8 of the 11 patients exhibiting undetectable HCV RNA within 12 weeks. HCV RNA reappeared in three patients (including two patients with SVC) exhibiting undetectable HCV RNA after 12 weeks. SVC was more frequent in patients with low viremia than in those with high viremia (55.6% vs. 14.3%; P=0.02), and in patients with HCV genotype non-1b than in those with HCV genotype 1b (57.1% vs. 22.2%; P=0.04). SVC was more common in patients with a > or =2 log reduction of HCV RNA at 4 weeks than in those with a smaller reduction (90% vs. 9.1%, P or =2 log reduction of HCV RNA at 4 weeks was a follow-up predictor for SVC. Undetectable HCV RNA occurring after 12 weeks was not sustained. All patients receiving antiviral therapy achieved a sustained viral response. Antiviral therapy should be initiated in patients with detectable HCV RNA at 12 weeks after the diagnosis.

Study on the relationship between T cell subsets and count and virus load in patients with hepatitis C / 安徽医科大学学报

Objective To investigate the correlation between T cell subsets and HCV viralload in HCV infection. Methods Flow cytometry was used to detect peripheral T cell subsets count of 69 patients with hepatitis C and 20 cases of normal health human, and fluorescent quantitative PCR was used to detect viral load in patients with hepa-titis C. Results ① The study showed that the percentage ratio of CD4 +T cells (42.87 ±6.11) and of CD4 +/CD8 +(1.34±0.25) in patients patients was significantly lower than those of normal health group,it was 49.55±6.68 and 1.82±0.11 (P<0.01);but the percentage ratio of CD8 +T cells (32.78±5.48) was higher than that of the normal health human ( 27.35 ±4.32 ) ( P<0.01 ) . There were no significant differences between the two groups in the percentage ratio of CD3 + T cells. ② The viral load gradually increased as the percentage ratio of CD8 +T cells increased, while the percentage ratio of CD4 +T cells and the percentage ratio of CD4 +T cells and CD4 +/ CD8 + were decreased gradually. Conclusion It may be one of the important reasons as the chronic infec-tion of hepatitis C virus by change of T cell subsets, and may be an important cause leading to the decrease of T lymphocyte response ability as the expression level of HCV RNA increases.

Comparison of COBAS AmpliPrep/COBAS TaqMan HCV Qualitative Test v2.0 with COBAS AMPLICOR Hepatitis C Virus Test v2.0 for the Qualitative Detection of Hepatitis C Virus RNA in Korean Clinical Samples / 임상검사와정도관리

BACKGROUND: We comparatively evaluated the performance of the conventional COBAS Amplicor HCV test v2.0 (CAM; Roche Molecular Systems, USA) and the newly developed COBAS AmpliPrep/COBAS TaqMan HCV test v2.0 (CAP/CTM; Roche Molecular Systems) for qualitative detection of hepatitis C virus (HCV) RNA in clinical samples. METHODS: Six hundred serum samples (100 HCV-positive, 500 HCV-negative, as determined by CAM) were selected and analysed using the new qualitative HCV RNA test, CAP/CTM qualitative test. Results were compared by confirmatory CAP/CTM quantitative test, which is a quantitative HCV RNA real-time polymerase chain reaction by Roche Molecular Systems, and anti-HCV test (Roche Diagnostics GmbH, Germany). Twenty-two additional serum samples, which gave a gray zone result by CAM, were selected for comparison. RESULTS: The two qualitative HCV RNA assays yielded concordant results for 586 of 600 tested samples (concordance rate, 97.7%; kappa coefficient, 0.92; 95% confidence interval [CI], 0.87 to 0.96; P<0.001). Upon re-testing by CAM, we found that the concordance rate increased to 98.2% (kappa coefficient, 0.93; 95% CI, 0.89 to 0.97; P<0.001). The additional 22 samples showing gray zone results for CAM were retested and were also tested by CAP/CTM. The results for 13 of these samples changed to negative and were now concordant with the CAP/CTM and confirmatory CAP/CTM quantitative results. For the remaining samples, the results were variable. For all the 22 samples, the results of the new CAP/CTM were in agreement with those obtained by confirmatory CAP/CTM quantitative test. CONCLUSIONS: The results of the two assays were in good agreement, with 97.7% concordance rate. However, CAP/CTM is more sensitive than CAM and showed no gray zone results. Therefore, it can be a more efficient and useful test for the qualitative detection of HCV RNA in clinical samples.