The Research Of Domestic Hepatitis C Antibody Testing Grey Area Set Up And Its Solutions

Objective: To set up clinical laboratory commonly used at present in Shanghai area four domestic anti- HCV antibody detection reagent grey zone, and testing results of each method grey area sample disposal scheme.Methods:(1) with rongsheng, the new wave, the progress and the four domestic anti-HCV detection reagent concentrations of standard, evaluation of the linear relationship of domestic kits and precision;(2) collection anti- HCV positive clinical test samples of 656 cases, respectively, with rongsheng, the new wave, the progress and the four domestic anti- HCV recheck, calculate different initial certification scope of review rate;(3) choose the anti- HCV inconsistent results in the early time of weakly positive samples of 338 cases with RIBA validation test and fluorescence quantitative PCR detection of HCV RNA as confirmation, RIBA and HCV RNA identified as the "gold standard", ROC curve drawing, discuss the optimal threshold, is about an index when the four largest domestic reagent best critical measurements/positive judgment value(S/CO value);(4) with 95% of the true positive and the negative of the S/CO value, analysis of fourdomestic anti- HCV positive detection reagent S/CO value Yin, grey zone; 36(5) analysis of four domestic reagent inconsistent results review anti- HCV and within the scope of the grey area, RIBA or HCV RNA positive samples, analysis of false negative rate;(6) analysis of four different domestic reagent under different concentrations of anti-HCV(S/CO < 1 and S/CO?1), HCV RNA detection rate and detection value;(7) four domestic anti HCV detection reagent in different anti- HCV concentration level relations with HCV RNA genes; Special in 12 cases(8) found that the "window period" sample, analyze antibody testing results, the relationship between RIBA confirm with HCV RNA, and genotyping.Results:(1) the standard concentration in 0.5 NCU /ml, four domestic detection system is not precision from 20.2%~36.2%respectively in the batch; Never precision from 18.9%~34.8%respectively. New wave and the linear relationship between the slope is very close to 2.495 and 2.510 respectively; Progress is relatively low as 1.348; Rongsheng minimum of 0.634.(2) four domestic anti- HCV detection reagent re-examination of negative rate were over 90%, S/CO value > 12.01, four domestic anti- HCV positive review detection reagent rate above 95%(3) rongsheng detection of anti- HCV best critical S/CO value of 1.31, the sensitivity was 94.8%, specificity of 94.6%, about an index is 0.894, the area under the ROC curve is 0.989; New wave detection of anti-HCV best critical S/CO value of 2.48, the sensitivity was 97.7%, specificity of 94.6%, about an index is 0.923, the area under the ROC curve is 0.989; Progress to detect anti-HCV critical S/CO value of 3.22, the best sensitivity was 97.2%, specificity of 97.3%, about an index is 0.945, the area under the ROC curve is 0.992; Families and the detection of anti- HCV best S/CO value of 4.32, the sensitivity was 96.2%, specificity of 94.6%, about index was 0.908, the area under the ROC curve is 0.991.(4) the four reagents are positive samples, RIBA and HCV RNA positive rate is as high as 96.3%(157/163); Inconsistent results and four reagent RIBA and HCV RNA positive rate 39.2%(31/79), four reagents are negative samples, RIBA and HCV RNA positive rate was 26.0%(25/96); 36 samples(5), the single reagent false negative rate was between 27.8%~94.4%, the two reagents after combination, the false negative rate was between 8.3%~61.1%;(6) four domestic testing results of S/CO<1 sample of HCV RNA detection rate was significantly lower than the S/CO?1sample detection rate; Anti- HCV S/CO<1sample of HCV RNA detection the median value were 3.62, 3.22, 3.31, 3.62, S/CO?1 of the median value of 6.67, 6.67, 6.65, 6.65(P < 0.05). In addition, the anti- S/CO?1each domestic reagent concentration of anti- HCV median are: 15.5, 49.6, 15.8, 49.6; And HCV RNA in a numerical all around 6.65.(7), 113 cases of HCV genotypes results, its genotype 1a 7(6.2%), 1b 72(63.7%), 2a 7(9.7%) and 3a(6.2%), 3b 11(9.7%), 6a 5(4.4%); In the S/CO<1and S/CO?1or genotype distribution under the different concentrations of no significant difference;(8) special samples of 12 cases for antibody detection results, four domestic test results were negative or part manufacturer in negative or positive grey zone, RIBA confirmation results for negative or uncertain, and HCV RNA viral load is weakly positive. Samples of 12 cases of RNA genotyping, 1b, 8 cases of early infection of genotyping may give priority to with 1b.Conclusion:(1) confirmed that four kinds of commonly used domestic anti- HCV antibody detection reagent best critical S/CO value, and for clinical provides four kinds of anti- HCV antibody detection reagent of grey zone(review);(2) the four kinds of anti-HCV antibody detection reagent solution, grey area results suggest progress, new wave, beauty can be used as reagents in rongsheng reinspection; Division China and the United States can be used as a new wave of reagent; Progress and beauty to each other both review reagents. When necessary, the double antigen sandwich method can be introduced to supplement the re-inspection.;(3) tries to analyze the causes of the grey area samples including analysis shows that in the early screening of the different way to detect the anti- HCV(ELISA, TRFIA, CLIA), the size of the sample S/CO value cannot represent the HCV RNA content; HCV genotypes with HCV levels had no significant correlation, no hint in the clinical diagnosis meaning; In serum HCV antibody detection at the same time, can increase the HCV RNA reagents for supplemental test, in order to reduce the residual window sample.