| BackgroudAlthough there is a risk of infecting pathogens in blood transfusion,sometimes leading to serious clinical consequences,blood transfusion is still an irreplaceable treatment in clinical practice at this stage,In order to ensure blood safety,the most critical step is to screen blood donors for blood transfusion and spread disease markers.Blood type,routine blood test,alanine aminotransferase(ALT),hepatitis B surface antigen(HBsAg),hepatitis C virus antibody(Anti-HCV),human immunodeficiency virus antigen and antibody(HIV Ag/Ab),Treponema pallidum antibody(Anti-TP)and nucleic acid test of HBV DNA,HCV RNA and HIV RNA(NAT)are the main tests for blood donors in China.NAT can detect the viral nucleic acids that appear earlier in blood samples.ELISA has long been used as a routine test for blood screening in blood stations.But there are disadvantages of high false positive rate and long window period.Nucleic acid detection is recognized as the best way to reduce the risk of missed detection and improve the detection rate of low viral load in ELISA window period.In 2015,the Ministry of health proposed the popularization of NTA screening HBV,HCV and HIV at national blood stations.Nucleic acid testing has been added to the enzyme-linked immunoassay for HBsAg,Anti-HCV,HIV/Ag/Ab and Anti-TP in blood donors.At present,most blood station laboratories use the 2 enzyme linked immunosorbent assay.Except for HBsAg,Anti-HCV,HIV Ag/Ab and Anti-TP double reagent positive samples,6/8 mixed nucleic acid detection was performed on the blood of double reagent negative and single reagent positive blood by ELISA.The mixed samples were positive,and then the nucleic acid detection mode was split.In this study,the unqualified rates of HBsAg,Anti-HCV,HIV Ag/Ab and Anti-TP in the blood samples of 151061 blood donors screened by Dezhou Central Blood Station from May 1,2015 to April 30,2018 were analyzed descriptively according to the distribution of different populations.Because the double ELISA positive blood is no longer tested for nucleic acid in the actual work of blood stations,we excluded the double ELISA positive samples for nucleic acid detection in the actual work and compared the detection results of HBsAg,Anti-HCV and HIV Ag/Ab kits from different manufacturers.The results of HBsAg screening tests with ELISA kits from different manufacturers were evaluated using NAT as gold standard.It will provide reference for blood stations to select screening models for markers of transfusion-transmitted diseases in the future.Objective1.Descriptive analysis of the population distribution of the unqualified rate of blood samples from unpaid blood donors.2.After eliminating ELISA double reagent positive samples,NAT test was carried out.To evaluate the screening effect of ELISA kits from different manufacturers on HBsAg in blood samples of ELIS A double-reagent positive donors,using NAT test results as gold standard.This study analyzed the screening effect of combined application of ELISA kits from different manufacturers.3.The role of nucleic acid detection as a supplement to HCV screening when used in conjunction with the third and fourth generation ELISA kits from different manufacturers.4.The role of nucleic acid testing as a supplement to HIV screening when used in conjunction with the third and fourth generation ELISA kits from different manufacturersMethodsResearch object Analysis of 150161 blood samples collected from blood donors in Dezhou in May 1,2015-2018 in April 30th.There were 113301 males and 35860 females in 150161 blood donors,aged 18-55 years.All blood donors were screened according to GB 18467-2011 Health Examination Standards for Blood Donors,which met the requirements of blood donors stipulated by the state.Experimental method ALT was detected by rate method;HBsAg,Anti-HCV,HIV Ag/Ab and Anti-TP were detected by enzyme-linked immunosorbent assay(ELISA);HBV DNA,HCV RNA and HIV RNA were screened by nucleic acid assay(PCR amplification technology)for double-reagent negative or single-reagent positive patients by ELISA.The blood station reagents are regularly tender annually,and the ELISA kit is not changed regularly.The sensitivity and specificity of ELISA test kits from different manufacturers were evaluated according to the results of preliminary re-examination of ELISA kits.Statistical method SPSS 24 software was used to analyze the data and evaluate the screening test.In statistical descriptions,chi-square tests using four-grid tables or R*C contingency tables were used to compare the rates of nonconformity of ALT,HBsAg,Anti-HCV,HIV/Ag and Anti-TP test results among different populations.At the same time,the chi-square test of four-grid table was used to compare the difference of evaluation index between different ELISA test kits and the difference of positive detection rate between ELISA kits and nucleic acid test.In addition,the chi-square test of R*C contingency table or four-grid table was used to compare the difference between the single positive rate of ELISA kits from different manufacturers and the positive rate of NAT test.Results1.Among the 151061 blood samples,there were 2324 unqualified blood samples,including 1815 of unqualified ALT,229 of positive Anti-HBsB,93 of positive Anti-HCV,179 of positive,Anti-TP,22 of positive HIV Ag/Ab,and the main causes of blood unqualified were unqualified ALT and Anti-HBsB positive.The unqualified rates of ALT,HBsAg,Anti-HCV,HIV Ag/Ab and Anti-TP in blood samples were significantly different among different groups,such as gender,age,occupation,education level and marital status.2.The screening effect of ELISA kit for HBsAg was analyzed after eliminating the positive double reagent test from different manufacturers,and the results of ELISA kit from different manufacturers and NAT test were compared.2.1 Analysis of screening effect of Xinchuang kit and Wantai kit for HBsAg positive after eliminating positive blood samples:A total of 90051 blood samples were tested by ELISA.After eliminating 115 positive blood samples,89936 ELISA-positive or ELISA-negative blood samples were tested for nucleic acid.89936 cases of blood samples were detected by NAT,and 45 cases of NAT positive blood samples were detected.The positive detection rate was 0.05%.Among them,there were 2 single-reagent positive blood samples detected by the Xinchuang kit,and no positive blood samples were detected by the Wantai kit in 45 NAT positive blood samples.Nucleic acid detection was used as the gold standard to evaluate the detection effect of two Xinchuang kits and Wantai HBsAg kit after eliminating positive blood samples.The sensitivity and specificity of the Xinchuang kit are 4.44%and 99.93%respectively.The sensitivity and specificity of Wantai kit were 0 and 99.91%respectively.The sensitivity and specificity of the series test in Xinchuang kit and Wantai kit were 0 and 100%respectively.The sensitivity of the parallel test in Xinchuang kit and Wantai kit is 4.44%and the specificity is 99.84%.2.2 Analysis of screening effect of Kehua kit and Rongsheng kit for HBsAg positive after eliminating positive blood samples:A total of 57312 blood samples were tested by ELISA.After eliminating 95 positive blood samples,57217 ELISA-positive or ELISA-negative blood samples were tested for nucleic acid.NAT was used to detect 57 217 blood samples,and 35 positive NAT blood samples were detected.The positive detection rate was 0.06%.In 35 positive NAT blood samples,neither Kehua kit nor Rongsheng kit detected single-reagent positive blood samples.Using nucleic acid detection as the gold standard,we evaluated the detection effect of Kehua and Rongsheng two kits on HBsAg after eliminating positive blood samples.The sensitivity and specificity of Kehua kit were 0 and 99.83%,respectively.The sensitivity and specificity of Rongsheng kit were sensitivity and specificity of the series test between Kehua test kit and Rongsheng kit 0 and 99.58%respectively.The were 0 and 100%respectively.The sensitivity of the parallel test is 0 and the specificity is 99.58%.2.3 The difference of positive rate of HBsAg in blood samples of donors was detected by ELISA kit and NAT after eliminating positive blood samples:In 90051 cases of blood samples,115 cases of double positive blood samples from X inchuang ELISA kit and Wantai ELISA kit were removed.64 cases(0.07%)were positive in the X inchuang kit,84 cases(0.09%)were positive in the Wantai kit,and 45 cases(0.05%)were positive in NAT.There was no significant difference in the positive rate of HBsAg among the three groups.In addition,there was no significant difference in the retest response rate between the Xinchuang kit and the Wantai kit.In 57312 blood samples,95 cases of double positive blood samples were removed from Kehua enzyme kit and Rongsheng enzyme kit.The difference of HBsAg positive rate among the three groups was statistically significant(?2=16.082,p<0.001).In addition,there was no significant difference in the retest response rate between the Kehua kit and Rongsheng kit.3.Comparison of the positive rate of Anti-HCV between the third and fourth generation ELISA kits and NAT in blood samples of blood donors after eliminating positive blood samples:In 104843 blood samples,the double positive reaction rate of the third generation Kehua and the fourth generation Wantai enzyme test kit was 59(0.06%).After eliminating the 59 blood samples,138 cases(0.13%)were found to be single positive with the third generation Kehua kit,and 41 cases(0.04%)were single positive with the fourth generation Wantai kit.No positive blood samples were detected in NAT.The single positive rate of the third generation Kehua kit was higher than that of the fourth generation Wan Tai Kit(?2=52.609,P<0.001).In addition,the retest reaction rate of the fourth generation Wantai kit was higher than that of the third generation Kehua kit(?2 = 6.301,p=0.012).In 45318 blood samples,23 cases had two positive responses to the Xinchuang and Wan Tai enzyme kit.After eliminating the 23 blood samples,39 cases(0.09%)were found to be single positive in the third generation Xinchuang kit,and 19 cases(0.04%)were single positive in the fourth generation Wantai kit.No positive blood samples were detected in NAT.The single positive rate of the third generation Xinchuang kit was higher than that of the fourth generation Wantai Kit(?2=6.901,p=0.009).In addition,there was no significant difference between the retest reaction rate of the fourth generation Wantai kit and that of the third generation Xinchuang kit.4.Comparison of the positive rates of HIV Ag/Ab in blood samples between the third and fourth generation ELISA kits and NAT kits after eliminating positive blood samples from different manufacturers:Of the 51248 blood samples,10(0.02%)were double positive in the third generation Rongsheng and the fourth generation Wantai ELISA kits.After eliminating these 10 blood samples,52 cases(0.10%)were detected by the third generation Rongsheng kit,49 cases(0.10%)by the fourth generation Wantai kit,and 1 case(0.002%)by NAT.The difference of positive reaction rate among the three was statistically significant(?2=38.275,p<0.001).It is worth noting that the 1 samples of NAT positive blood samples were detected by the fourth generation Wantai kit.In addition,the third generation Rongsheng's retest rate was higher than that of the fourth generation Wantai kit(?2=8.255,p=0.004).In 98913 blood samples,the double positive reaction rate of the third generation Kehua and the fourth generation Wan Tai enzyme test kit was 12(0.01%).After eliminating the 12 blood samples,23 cases(0.02%)were found to be single positive with the third generation Kehua kit and 101 cases(0.10%)were detected with the fourth generation Wantai kit.No positive blood samples were detected in NAT.The single positive rate of the fourth generation Wantai kit is higher than that of the third generation Kehua kit(?2=49.095,p<0.001).In addition,there was no significant difference in the rate of retest reaction between the third generation Kehua kit and the fourth generation Wantai kit.Conclusion1.The unqualified rate of blood samples of unpaid blood donors varies in different populations.2.The results of ELISA and NAT were inconsistent after screening out positive double reagents from different manufacturers.The NAT test of HBV can find that 0.05%?0.06%of the single positive or double negative blood samples detected by ELISA are not qualified.The current NAT detection mode is of great significance for the screening of HBV.3.After screening the positive results of two ELISA kits from two different manufacturers,the detection rate of HCV by mixed NAT at present as a supplementary test is zero.4.After screening the positive results of two ELISA kits from two different manufacturers,only one of the 150 161 cases of HIV screening by mixed NAT at present as a supplementary test was detected,and this case was positive by single reagent of Wantai fourth generation reagent. |
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