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Objective To reveal the characteristics of S gene sequence of hepatitis B surface antigen (HBsAg) in hepatitis B virus (HBV)-infected patients with low HBsAg level.Methods From February 2016 to December 2017, 1 308 serum samples of inactive HBsAg carriers were collected from the 903rd Hospital of PLA and Hangzhou Jianggan District People′s Hospital.The cases were divided into high-level group and low-level group according to the level of serum HBsAg (10 IU／mL) expression.The HBV S gene was sequenced in patients with low-level HBsAg expression.In addition, in patients with high-level HBsAg, 100 patients were randomly selected (stratified sampling) for HBV S gene sequencing based on the matching of age and serological pattern (hepatitis B e antigen [HBeAg] negative) of low-level HBsAg group.A comparative analysis was conducted between HBV S gene sequences from inactive HBsAg carrier in low HBsAg expression group and the HBV reference S gene sequences from inactive HBsAg carrier in high HBsAg expression group .The results of normal distribution data were expressed as Mean ±SD, and analyzed using t-test.The results of non-normal distribution data were expressed by M(QR), and analyzed using Mann-Whitney U test.Chi-square test or Fisher exact test was used to compare continuous variables and classification variables between the two groups .Results There were 276 serum samples from the low level group and 1 032 serum samples from the high level group , including 257 HBsAg／HBeAg／anti-HBc-positive cases, 753 HBsAg／anti-HBe／anti-HBc-positive cases, and 22 HBsAg／anti-HBc-positive cases.Successful HBV S gene sequencing was performed on 126 out of 276 patients in the low-level HBsAg group.According to the age inthe low-level HBsAg group, 100 samples with negative HBeAg in the high-level HBsAg group were randomly selected , among which 94 patients were genotyped and hemotyped.The results showed that there were statistically significant differences in HBV serological markers , HBV DNA level and HBV genotype distribution between the high level group (94 cases) and the low level group (126 cases) (all P＜0.05).The ASC-R-B and ASC-R-C genotypes reported in this study had high homology (99.6%-100.0%) with those reported in Shanghai , Chengdu, Wuhan, Yunnan and Beijing of China , and high homology (98.2%-99.6%) with those reported in Japan and Korea of NCBI genotype B and C reference sequences, but had low homology with patients far away from China (98.2% in Canada and 98.7% in Indonesia).In genotype B of the low level group , the amino acid mutation number of SHB protein was 71, and the hot spot mutation number was 19, both higher than those in the high level group (39 and 8, respectively). The difference was statistically significant (χ2 ＝12.303 and 4.766, respectively, both P＜0.05).Amino acid mutation sites in the low HBsAg group were mainly distributed on both sides of the major hydrophilic region (MHR) (amino acid residues 40 -49 and 198 -220).There were no significant differences in amino acid mutation number and hot spot mutation number between the two groups of C genotype (χ2 ＝0.383 and 0.409, respectively, both P＞0.05).For genotype B, 12 single point mutations and 4 dual co-mutations were found in low level group.Among them, one single point mutation (S210R) and 3 dual co-mutations (G44E／V+T45P／I, G44E／V+L49P／R and N40S+I208T) were not hot spot mutations , while 2 dual co-mutations and 2 single point mutations were found in high level group.The difference between two groups was statistical significant (χ2 ＝7.533,P ＝0.006).For genotype C, 5 single point mutations ( T5A, A45T, T47A／K, Q101R and I126S／T) were found in low level group and 1 single point mutation (N3S) in high level group.The difference in mutation frequency between two groups were statistical significant (χ2 ＝47.914,P＝0.000).Conclusions Significant mutations in multiple regions and at multiple sites ( including co-mutations) on both sides of the MHR may be one of the causes of low HBsAg expression level in this population .
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Objective To investigate the changes of HBV markers in patients with HBeAg positive chronic hepatitis B(CHB) after a large number of blood transfusion treatment.Methods 26 patients with chronic HBV that were treated massive blood transfusion in 24h were collected from Jan 1,2015 to Jan 1,2016.The HBV serum markers and HBV-DNA were measured and compared before and after treatment.Results The HBsAg,anti-HBs and anti-HBc concentration in first day after treatment were different compared with before treatment(t=2.681,4.753 and 5.116,all P<0.01).The HBsAg,anti-HBs and anti-HBc concentration in third day after treatment exist differences compared with before treatment(t=1.681, 2.209 and 3.118,all P<0.05).The difference still exist in the seventh day after treatment compared with before treatment with only anti-HBc concentration(t=2.463,P<0.05).There was not difference of HBeAg and HBV-DNA before and after blood transfusion in patients(t=0~1.132,P>0.05).After transfusionthe concentration of HBsAg in the fifth day was the lowest concentration as 0.17±0.03 IU/ml,the seventh day rose to 387.50±31.89 IU/ml,reaching the highest value,and the concentration of HBsAb decreased gradually to minimum at the seventh day that was 1.51±5.98 mIU/mmol,and the concentrations of HBeAg,HBeAb and HBcAb had no obvious change.Conclusion The HBsAg,anti-HBs and anti-HBc could be changed in patients with HBeAg positive CHB after massive transfusion therapy in short term.HBeAg and HBV-DNA were not affected by transfusion therapy.
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ObjectiveTo discuss the clinical significance of han nationality and kazak hepatitis B virus large surface protein detection.MethodsEnzyme linked immunosorbent assay(ELISA) method was used to examine the HBV-LP、HBV markers and quantitative real-time PCR methods were used to detect the HBV DNA in 270 patients with Hepatitis B.ResultsAmong the 270 cases,there was no significant difference between the levels of HBV DNA and HBV-LP( P ＞ 0.05 )in HBe Ag-positive patients,which was not affected by nationality.Significant difference of positive rate was observed between HBV-LP and HBV DNA( P ＜0.05) in HBe Ag-negative ones,which was not affected by nationality.HBV-Lp expression was significantly correlated with the logarithm of HBV DNA level ( r =0.986,P ＜ 0.05).ConclusionThere was higher coincidence rate between the levels of HBV-LP and HBV DNA in HBeAg--positive patients.The positive rate of HBV-LP was higher than that of HBV DNA in HBe Ag-negative patients.HBV-LP could serve as a reliable marker in the reflection of HBV the replication at protein level,and it was valuable to monitor HBV replication and prognosis of the disease,especially in HBe Ag-negative HBV infected patients.
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OBJECTIVE To investigate the correlations on hepatitis B virus(HBV)preS1-antigen(preS1-Ag)and HBV-DNA,HBV markers in patients with hepatitis B.METHODS The markers,preS1-Ag and HBV-DNA were determined by ELISA and PCR in 406 patients with hepatitis B and 80 healthy persons.RESULTS The positive rates of preS1-Ag in 160 patients with HBsAg(+)and HBeAg(+)were 84.4%.The positive rates of preS1-Ag in 246 patients with HBsAg(+)and HBeAg(-)were 42.3%;the difference between them was significant(?2=70.9,P
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OBJECTIVE To investigate the relationship among hepatitis B virus(HBV) basic core promoter(BCP) mutation,serum HBV DNA contents and HBV markers as well as liver function of chronic hepatitis B patients.METHODS The A to T mutation at nucleotide 1762 and G to A mutation at nucleotide 1764 were determined by the method of polymerase chain reaction(PCR) microplate hybridization ELISA in 176 patients with chronic hepatitis B virus infection.RESULTS The rate of HBV BCP mutants in negative and positive groups of HBeAg was 49.4% and 33.3%,respectively(P
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OBJECTIVE To study the relativity among PreS1-Antigen,HBV markers and HBV-DNA. METHODS The HBV markers,PreS1-antigen and HBV-DNA were determined by ELISA and PCR in 102 patients with chronic hepatitis B and 73 healthy persons. RESULTS Among 102 patients with chronic hepatitis B,the concordance rate of PreS1-antigen and HBeAg with HBV-DNA was 70.6% and 75.5%.The sensitivity of PreS1 was better than HBeAg but the specificity was contrary.It represented some patients with HBeAg(-) still had viral replication.On the increase in the level of HBV-DNA,the positive rates of PreS1-antigen and HBV markers increased. CONCLUSIONS The detection of PreS1-antigen can well reflect the HBV replication.The synchronous dynamic detection of PreS1-Antigen,HBV markers and HBV-DNA has its important clinical meaning.