Comparative Analysis On The Results Of Serum HBsAg And HBeAg And HBV-DNA With Different Methods In Different Laboratories

Objective:To compare the consistency and variability of the results of HBsAg and HBeAg detected with difference methods of TRFIA,MEIA and ECL and of the results of HBV-DNA detected using fluorescence quantitative PCR in different labs,the serum specimens of 90 cases of chronic hepatitis B patients were studied.And at the same time,the correlations among HBsAg,HBeAg and HBV-DNA in every lab were analyzed.Methods:The serum specimens of 90 cases of chronic hepatitis B patients were collected.HBsAg and HBeAg were tested separatedly by TRFIA,MEIA and ECL in 4 labs,and HBV-DNA was tested by fluorescence quantitative PCR in corresponding labs.Results:①Serum specimens were tested in four laboratories.The total coincidence was 100%and 85.6%in HBsAg and HBeAg respetively.MEIA as the golden standard,it had a better consistency between TRFIA(lab1)and MEIA(lab3)and that of between ECL(lab4)and MEIA(lab3),and the former(Kappa=0.914)was higher than the latter(Kappa=0.905).ECL as the golden standard,it had a better consistency(Kappa=0.821)between TRFIA(lab 1)and ECL(lab 4).On the detecting of the positive rate in HBeAg,there were significant differences(p＜0.05)between TRFIA(lab1)and TRFIA(lab2),between TRFIA(lab1)and ECL as well as between TRFIA(lab2)and MEIA.However,there was no significant differences between MEIA and ECL(p＞0.05).On the quantitative detection of HBeAg by using TRFIA, there was no significant differences(p＞0.05)between lab1 and lab2.In HBeAg qualitative judgement,13 serum specimens had relatively inconsistent,of which two cases may be false negative and seven cases may be false positive.②On the quantitative detection of HBV-DNA by using fluorescence quantitative PCR,103 copies/ml as the boundary point,the total coincidence was 83.3%,the consistent ratios of HBV-DNA positive results were 88.4%and 94.1%between lab2 and lab3 and between lab3 and lab4 respectively and lower than that among other laboratories.The ratio of less than±1 log took on an average of 79.3%in the differences in HBV-DNA level among laboratories, the difference was not significant through statistical analysis(p＞0.05).On the positive rate in HBV-DNA,there was no significant difference between lab1 and lab4 and between lab2 and lab3.In HBV-DNA quantitative judgement,16 serum specimens were relatively inconsistent, of which there were five false negative cases,five false positive cases and four unreliable cases.③Using TRFIA for detecting HBsAg,HBeAg and fluorescence quantitative PCR(lab1)for detecting HBV-DNA,the results showed that the correlations between HBsAg and HBeAg, and between HBsAg and HBV-DNA existed as well as HBeAg and HBV-DNA(p＜0.05)when HBeAg was positive.However,there was no correlation between HBsAg and HBeAg and between HBsAg and HBV-DNA when HBsAg was less than 225ng/ml.In addition,there was no correlation between HBsAg and HBV-DNA when HBeAg was negative(p＞0.05).Using TRFIA(which was different with lab1 in instruments)for detecting HBsAg,HBeAg and fluorescence quantitative PCR(lab2)for detecting HBV-DNA,the results showed that the correlations between HBsAg and HBeAg,and between HBsAg and HBV-DNA existed as well as HBeAg and HBV-DNA(p＜0.05)when HBeAg was positive.in addition,there is no correlation between HBsAg and HBV-DNA when HBeAg was negative(p＞0.05).Using ECL for detecting HBsAg,HBeAg and fluorescence quantitative PCR(lab4)for detecting HBV-DNA,the results showed that the correlation between HBsAg and HBeAg,and between HBsAg and HBV-DNA existed(p＜0.001)when HBeAg was positive.But there was no correlation between HBsAg and HBV-DNA when HBeAg was negative(p＞0.05).Conclusion:①Regardless of using any reagents and methods,there was a high degree consistency in positive results of serum HBsAg in patients with chronic hepatitis B in different laboratories(100%in this study).However,there were differences in the positive rates of HBeAg among different methods and laboratories,the overall difference was up to 14.4%(13/90).To specimens,especially close to the positive(or negative)critical value, retesting on the specimen and verifying among different laboratories were recommended clinically.②There was also some difference in testing HBV-DNA in different laboratories and the overall difference was up to 17%(16/90).However,there was a high degree consistency in the compared results in different laboratories with each other and the laboratories which had high degree consistency could use different instruments and reagents.It suggested that the normative operation and stability of the techniques had more effects on the results of testing HBV-DNA with fluorescence quantitative PCR.③Recently,quantitative detection of HBsAg and HBeAg is a hot issue clinically.This study showed that the quantitativeness in HBsAg and serum HBV-DNA level was positively correlated as well as quantitativeness in HBeAg and serum HBV-DNA level when HBeAg was positive in a certain range.It suggested that in addition to quantitative detection on HBV-DNA,the quantitative detection on HBsAg and HBeAg could reflect HBV-DNA level and fluctuation condition and the response to treatment in patients with chronic hepatitis B to a certain extent.However,HBsAg could not reflect HBV-DNA level as HBeAg was negative. In addition,in this study the number of samples was smaller in HBeAg-negative patients with chronic HBV infection.So it is necessary to enlarge the sample of HBeAg-negtive patients with chronic HBV infection to clarify the relationship between HBsAg and HBV-DNA.