Detecting the occult HBV infection: a laboratory study / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To see the HBV DNA detection instance in the HBsAg negative people and to study the serological method detection strategy for detecting hepatitis B virus large surface protein (HBLP) to filtrate the occult HBV infection.</p><p><b>METHODS</b>The HBsAg negative serum samples were divided into HBsAb negative and positive two species according to the hepatitis B virus markers (HBVM) in daily work excepting the special HBVM modes. Total 2000 stochastic serum samples with 1000 HBsAb negative results and 1000 HBsAb positive results were collected from the copy tubes to detect HBVM with national ELISA reagent kits and put them -20 degrees C frostily. Mixed samples (8 x 30 microl) were analyzed with fluorescence quantitative PCR (FQ-PCR) and filtrated the individual positive samples. The filtrated samples were doubly tested again with American MONOLISA HBsAg ULTRA reagents.</p><p><b>RESULTS</b>No HBV DNA positive results were found out from the 1000 HBsAb positive samples and 19 cases HBV DNA positive results were found out from the 1000 HBsAb negative samples. On these 19 samples, the HBsAg results from the American MONOLISA HBsAg ULTRA reagents were all positive and the HBLP results were all positive, too. The 19 HBV DNA quantitative results were divided into 2 cases more than 500 copies/ml, 3 cases between 400-500 copies/ ml, 3 cases between 300-400 copies/ml, 7 cases between 200-300 copies/ml and 4 cases between 100-200 copies/ml.</p><p><b>CONCLUSION</b>The leaked samples tested HBsAg with national reagents are mostly from the HBsAb negative people. HBLP results may be positive on these samples and detecting HBLP marker is propitious to filtrate the occult HBV infection. This study provided a kind of serological reference for actively searching for the detecting strategy in occult HBV infection field.</p>

Detecting hepatitis B virus large surface protein to filtrate the occult HBV infection / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Detecting hepatitis B virus large surface protein (HBLP) with serological method to filtrate the occult HBV infection and study the clinical detection strategy.</p><p><b>METHODS</b>Two thousands HBsAg negative stochastic serum samples were collected from the copy tubes in daily work to detect hepatitis B Virus markers (HBVM) with national EHSA regent kits and put them -20 degrees C frostily. The 2000 samples were detected with HBLP and filtrated the positive samples. The HBsAg markers were doubly counterchecked with other two adding kinds of national ELISA regent kits (total 3 kinds) at the filtrated samples. The last samples were doubly tested again with American MONOLISA HBsAg ULTRA regents. HBV DNA levels were quantitative analyzed with fluorescence quantitative PCR (FQ-PCR) and taking the mean results.</p><p><b>RESULTS</b>Fifteen HBLP positive samples were detected out from the 2000 serum samples. We educed the conform negative results from the three kinds of national regents but conform positive results from the American MONOLISA HBsAg ULTRA regents in repeating HBsAg detection at the 15 samples. The HBV DNA FQ-PCR quantitative results were all less than 500 copies/mi and divided into two cases hetween 400-500 copies/nil, three cases 300-400 copies/ml, five cases 200-300 copies/ml, four cases 100-200 copies/ml and one case was less than l00copies/ml.</p><p><b>CONCLUSION</b>The false HBsAg negative results for serum samples are more generally through national regents than through importations and HBLP results mayhe are positive in these samples. Detecting HBLP marker is propitious to filtrate the occult HBV infection. This study provided a kind of serological reference for actively searching for the detecting strategy in occult HBV infection field.</p>

Expression,purification and identification of hepatitis E virus pb166-GST fusion protein / 中华检验医学杂志

Objective To express,purify and identify recombinant hepatitis E virus(HEV) pb166-GST fusion protein using GST gene fusion system and investigate its potential role in researching Hepatitis E diagnostic antigen field.Methods The recombinant E.coli BL21 performed by our own laboratory was used to induce the HEV pb166-GST expression with IPTG.The products were purified by BD Biosience GST purifying system.The specific expression was identified by SDS-PAGE and Western blot.The experiment conditions and results were described and analysed.Results The resolved HEV pb166-GST fusion protein on SDS-PAGE showed a major band at position of 43 kD.The expressed proteins had a single expected band after purify and the protein was recognized by anti-GST antibody on PVDF membrane.Conclusion The recombinant HEV pb166-GST fusion protein is expressed in recombinant E.coli BL21 efficiently in this way,and might be used as a candidate for diagnostic antigen of HEV.