Application Of Hepatitis B Virus Large Surface Protein As A Marker In Hepatitis B Detection

ObjectiveAt present,there are about 350 million chronic HBV carriers in the world,and the positive rate of hepatitis B vires surface antigen(HBsAg) is about 9.75%in China. Nowadays,there are more than 20 million patients with hepatitis B in China.About 270,000 people(23/10 million) die from hepatitis B-related diseases annually. Therefore,it is very important to establish a simple,sensitive and reproducible method for HBV detection.Yet the current detection assays for HBV replication in clinical laboratory have different clinical significances.Some of them are short of specificity, and some are difficult to be used in basic hospitals.The anti-HBV drugs have some therapeutic efficacy,but no reasonable and effective detection index to evaluate therapeutic effective has been found,and the time when the anti-HBV drugs should be stopped is still not determined.In addition,the index to signify whether recidivation could occur or not after drug withdrawal is not found.Therefore,the laboratory index is needed badly to be found,which could effectively evaluate and signify viral replication and discovers hepatitis B patients in active stage from HBeAg-negative asymptomatic carriers.With the rapid development of molecular biology,immunology and computer technique and so on,a new laboratory index:Hepatitis B Virus Large Surface Protein (LHBs) has been used gradually in laboratory diagnosis.However,no mature detection of LHBs was used in clinical laboratory detection up to date.By detecting a great number of clinical samples,the present study verifies that LHBs is of specificity in HBV replication of hepatitis B patients.The aim of this study is to lay a foundation in both theory and experiment for the improvement of HBV prevention and cure in our country.Methods1.Clinical specimen collection:The sterile venous blood of patients was collected with blood collection needle.2.First of all,the patients who were infected with HBV were determined by detecting Hepatitis B serum makers.3.Detecting the number of virus replication of above mentioned patient serum by PCR assay.4.Detecting LHBs in the above serum samples through ELISA.5.Evaluating the concordance between the detecting results of LHBs and PCR assay of HBV by detecting a large number of clinical samples.Results1.The protein level of LHBs and the logarithm of HBV DNA copy number have good dependablity,the correlation coefficient(r)=0.932.Variance analysis showed that LHBs content was significantly different between different groups of HBV DNA copies.2.By chi-square test,the difference between the positive rate of LHBs and HBV DNA is not significant among hepatitis B patients with HBsAg,HBeAg and anti-HBc (P＞0.05).3.The positive detection rate of HBV DNA and LHBs from hepatitis B patients with HBsAg,anti-HBc and anti-HBe are 51.12%(181/354) and 55.65%(197/354), respectively.There was no significant difference between two assays by chi-square test (X~2=0.81,P＞0.05);4.Five hundred and ninety-three sera from HBsAg-positive patients were tested by the laboratory index of HBV DNA and LHBs,respectively.The results show that there is no significant difference between two assays(P＜0.005),and the results of this two assays are identical according to benchmark of a=0.05.In summary,there is no significant difference between the results of LHBs detection and HBV DNA detection.The logarithm value of HBV DNA copies is consistent with expression of LHBs(r=0.932,P＜0.001),the change of logarithm value of HBV DNA copies can be explained by linear regression model with LHBs OD value as variable.Conclusions1.LHBs can reflect HBV replication,and there is a good correlation between LHBs and HBV DNA copies in sera.2.LHBs is new serological immunological indicator for reflecting the HBV replication of HBeAg-negative patients,which lay an experimental basis for clinical detection of HBV replication.3.A reasonable laboratory detecting assay to determine the therapy end-point of hepatitis B is found.