HBsAg Confirmatory Test And The Application And Analysis Of Serum MiRNA-122and Other Indexes In CHB Patients

Objective To explore antibody neutraliza tion confirmatory test in newly diagnosed HBsAg positive patient; and to observe serum miRNA-122levels during four different phases with hepatitis B virus (HBV) infection; and to study the changes and significance of HBV related indexes in chronic hepatitis B virus (CHB) patients with serum HBV-DNA low replication index. Method To collect1974positive samples tested by chemiluminesent micropaticle immunoassay (CLIA) and re-examine the HBsAg by gold-labeled immunoassay (GICA), enzyme-linked immunoasorbent assay(ELISA) and other methods for statistical analysis. Meanwhile, the HBsAg CLIA positive but HBsAg ELISA negative samples were further confirmed by CLIA neutralization confirmatory test. MiRNA-122, HBV-DNA, HbeAg, HbsAg and ALT levels were mainly detected with real time fluorescence quantitative PCR (FQ-PCR) and CLIA in80CHB patients. The results were compared and analyzed with linear correlation analysis.229cases of blood specimens from patients with HBV-DNA low copy, ALT, HBV markers, mIL-2R, IL-10, miRNA-122and other indicators are using automatic analyzer, and compared to the linear correlation analysis on the results.Results The HBsAg CLIA positive samples were re-examined by GICA and ELISA. There were undetected situation in GICA and ELISA methods. The missed detection rate was respectively9.1%(180/1974) and25.4%(501/1974) in HBsAg-positive samples. These undetected samples were mainly in the HBsAg CLIA negative samples of which the test value was lower than10.0IU/ml. And the concentration range accounted for as high as26.90%(531/1974) in HBsAg negative samples.180HBsAg CLIA positive samples which undeteced by ELISA were confirmed by CLIA neutralization test and the positive rate was96.67%(174/180).6cases which could not be confirmed were tested by HBV-DNA detection and two weeks’ follow-up.3cases were confirmed and the other3were ruled out. MiRNA-122levels were obviously high during four different phases, especially in immune tolerant phase and in immune clearance phase, showing a significant difference (P<0.01).Meanwhile, there was a significant linear correlation between miRNA-122levels and HbeAg and HBV-DNA levels (r=0.263, r=0.171).But there was a significant linear correlation between miRNA-122levels and HbsAg and ALT levels (r=0.597, r=0.767). There were38%of HBeAg positive among patients with HBV-DNA low copy. HBeAb positive patients accounted for51.53%. In229cases of patients, ALT elevated was66.81%of the total. The levels of serum miRNA-122was significantly correlated with ALT(r=0.841). The concentration of IL-10and mIL-2R was significantly different (P<0.05) between HBeAg positive-and-negitive group and control group. Conclusion Compared with quantitative methods as CLIA, the qualitative methods such as GICA and ELISA were obviously insufficient. It was so important to detect the low-concentration HBsAg-positive samples because it helped HBsAg-positive patients to early detect, early diagnose and early treat.The laboratory should prefer sensitive CLIA for the first-time HBsAg detection and collect double serum sample for the low-concentration HBsAg-positive patients. The confirmatory tests were done by HBV-DNA detection and follow up to create the file. It avoided misdiagnosis and missed diagnosis. Maybe there had no relation between miRNA-122and the viral replication. But miRNA-122was still important to improve the diagnosis, assessment and health management of patients with HBV infection. In patients with HBV-DNA low copy, the changes of ALT, mIL-2R, IL-10, miRNA-122and other indicators proved substantive liver cell damage. Clinical attention should be paid to. Especially, it could not be generalized with non-active HBV carrier state.