ABSTRACT
Objective@#To investigate the comparative study of serum hepatitis B virus (HBV) large protein (HBV-LP) , HBV-DNA, and Pre S1 antigen (Pre S1-Ag) detection in the evaluation of serum HBV replication in patients with chronic hepatitis B.@*Methods@#A total of 482 patients infected with chronic hepatitis B virus (CHB) were enrolled and the serums were collected in a hospital of Hefei city in Anhui province from June 2013 to March 2015. The serum HBV-LP, HBV markers(HBV-M) and Pre S1-Ag were detected using ELISA, and HBV-DNA were quantified using quantitative real-time PCR. The positive detection rate difference of HBV-DNA, HBV-LP and Pre S1-Ag were compared, the correlation between the logarithm of HBV-DNA copies number and the absorbance value of HBV-LP was analyzed using Spearman rank correlation.@*Results@#The positive rates of HBV DNA, HBV-LP, and Pre S1-Ag were 67.22% (324/482), 73.86% (356/482), and 37.34% (180/482), respectively (P<0.01). The positive rates of the three markers were 54.57% (185/339), 64.90% (220/339), and 27.73% (94/339), respectively, in 339 HBeAg-negative CHB patients (P<0.01). In HBeAg negative patients, the positive rate of HBV-LP in 185 cases of HBV-DNA positive samples was 90.81% (168/185), which was higher than that of Pre S1-Ag with rate of 39.46% (73/185) (P<0.01).The positive rate of HBV-LP was 33.77% (52/154) in 154 cases of patients with negative HBV-DNA whose positive rate was higher than Pre S1-Ag with positive rate of 13.64% (21/154)(P<0.01). The positive rates of HBV-DNA, HBV-LP and Pre S1-Ag in HBsAg, HBeAg and HBcAb positive groups were 97.04% (131/135), 94.81% (128/135), and 60.00% (81/135), (P<0.01); The positive rates of three indexes of HBsAg, HBeAb, HBcAb positive group were 53.74% (122/227), 63.88% (145/227), and 27.31% (62/227); The positive rates of three indexes of HBsAg and HBcAb positive group were 55.79% (53/95), 67.37% (64/95), and 28.42% (27/95) (P<0.01). The absorbance value of HBV-LP was positively related with the logarithm of HBV-DNA copies number (Spearman rank correlation coefficient was 0.908, P<0.01).@*Conclusion@#There was a good correlation between HBV-LP and HBV-DNA load value, and could be used as an effective complement for the detection of HBV-DNA and HBV-M. Compared with Pre S1-Ag.
ABSTRACT
Objective To investigate the relationship between Pre-S1 antigen and hepatic function of the patients with chronic hepatitis B.Methods Serum samples of 300 cases with chronic hepatitis B patients were collected,Pre-S1 antigen and alanine aminotransferase (ALT),aspartate aminotransferase (AST) were checked,and the correlation were analyzed.Results Among 300 cases,patients with positive Pre-S1 antigen were 58.33％(175/300).In patients with positive Pre-S1 antigen,the abnormal ALT(＞40 U/L) detection rate was 78.75％ (138/175),the abnormal AST(＞40 U/L) detection rate was 77.17％ (136/175),Pre-S1 antigen negative patients ALT abnormal detection rate was 51.20％ (64/125),AST anomaly detection rate was 42.40％ (53/125).There was significant difference between two groups(x2 =2.875,5.223;P＜0.05).Conclusion In patients with chronic hepatitis B,Pre-S1 antigens can be used as a marker of hepatitis B virus infection,copy and hepatic cell injury.
ABSTRACT
Objective To investigate the clinical significance of combned detection of serum pre-S1 ,pre-S2 antigen and hepatitis B virus(HBV) e antigen(HBeAg) for diagnosis of HBV infection .Methods Enzyme-linked immunosorbent assay(ELISA) was employed to detect the serum pre-S1 ,pre-S2 antigen and HBeAg of 450 patients with chronic hepatitis B and real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect their HBV DNA .Results In patients with different HBV DNA levels ,serum pre-S1 ,pre-S2 antigen and HBeAg showed different expression ,the higher the load ,the higher the expression (P<0 .05) .Pre-S1 ,pre-S2 antigen and HBeAg were all correlated with HBV DNA (P<0 .05) and the descending order of rele-vance were HBeAg ,pre-S1 and pre-S2 antigen .Combied detection of the three indicators showed the highest correlation with HBV DNA .The positive proportion of HBV DNA of patients with the three indicators positive was markedly higher than those in pa-tients with single or two indicators positive (P<0 .05) .Conclusion Combind detection of serum pre-S1 ,pre-S2 antigen and HBeAg may reflect well on HBV replication .
ABSTRACT
Objective: To study the correlation between HBV Pre-S1 antigen, HBeAg levels and HBV DNA copies, so as to assess the clinical value of Pre-S1 in detection of HBV replication. Methods: A total of 363 HBsAg-positive samples were collected. The levels of Pre-S1 antigen, HBeAg and HBV DNA copies were determined by ELISA, time-resolved immuno-fluorescent method and real-time fluorescent quantitative PCR(FQ-PCR),respectively. The correlation between the determination results was analyzed. Results: Pre-S1 antigen level was correlated with the level of HBeAg (χ2 = 94.4,P<0.01), with the coefficient being 0.45; Pre-S1 antigen level was also correlated with HBV DNA(χ2 = 198.58, P<0.01), with the coefficient being 0.59. The positive rate of Pre-S1 increased with HBV DNA copies. When logarithm of HBV DNA copies was more than 3, the sensitivity, specificity, accuracy, positive predictive value(PPV), positive likelihood ratio(PLR) and odds ratio(OR) of Pre-S1 were 84.5%,91.2%,87.0%,94.1%,9.6,and 56.8 respective; the numbers of HBeAg were 50.4%,94.9%,67.2%,94.2%, 9.8 and 18.9, respectively. Conclusion: Pre-S1 antigen is correlated with HBV DNA and is more sensitive and accurate than HBeAg in reflecting HBV replication. The above 3 parameters can complement each other and the combination of them is more accurate in clinical diagnosis.
ABSTRACT
Objective To evaluate the value of pre-S1 Ag in combined detection of hepatitis B virus. Methods 288 patients' serums with chronicity type B hepatitis (CHB)were collected when they had been detected with HBV-DNA,HBVM or ALT/AST,and then ELISA assay were applied to detect pre-S1Ag. Results A total of 288 patients ,including 175 males and 113 females( their meso-age is 36.89 years old) were recruited into this study. 161 patients were pre-S1 Ag positive(68.4%), 197 patients' HBV DNA contents exceeded 5.0E + 2 copy/ml(55.9%) ,71patients were H BeAg positive (24.6%), 186 patients were HBeAg negative (64.58%). The rate of 64 patients' ALT and AST exceeded 0. 84, and their' ALT exceeded 40 u/ml ( 22. 2% ). 60 patients' AST exceeded 50 u/ml (20. 8% ). These patients were divided into pre-S1 Ag positive group and pre-S1 Ag group. In pre-S1 Ag positive group, 124 patients' HBV DNA contents exceeded 5.0E + 2 copy/ml( 62.9% ), 118 patients were HBeAg negative (59.8%). When HBV DNA content was less than 5.0E+2 copy/ml,pre-S1 Ag positive rate was 35.2% and pre-S1 Ag negative rate was 63.6%. when HBV DNA content was less than 5.0E + 3 copy/ml, pre-S1 Ag positive rate was 56.9% and pre-S1 Ag negative rate was 81.7%. When HBV DNA content exceeded 5.0E + 5 copy/ml but less than 5.0E + 8 copy/ml ,pre-S1 Ag positive rate was 33% and pre-S1 Ag negative rate was 14.3% ,so two groups had significant difference. Conclusion Pre-S1 Ag positive and HBeAg negative in the CHB patients serums signify virus replication,and it can help us decide prognosis.
ABSTRACT
This study investigated the prevalence of the precore G1896A mutation in Chinese patients with hepatitis B e antigen (HBeAg) negative HBV infection and its relation to serum HBV pre-S1 antigen. The overall prevalence of the precore G1896A mutation was 72.6 percent in HBeAg-negative Chinese patients with detectable serum HBV DNA. The prevalence of the precore G1896A is significantly higher in Chinese HBeAg-negative patients with chronic hepatitis B than that in inactive HBV carriers with detectable serum HBV DNA. Serum pre-S1 and the precore G1896A mutation were simultaneously detected in most of Chinese HBeAg-negative patients.
Subject(s)
Humans , Hepatitis B virus , Hepatitis B, Chronic , Mutation , Prevalence , Virus Diseases , Diagnostic Techniques and Procedures , Methods , MethodsABSTRACT
Objective To discuss the value of detection of Pre S1 antigen(Pre-S1 Ag) of hepatitis B virus. Methods ELISA was used to detect Pre-S1 Ag and HBeAg in serum from 854 HBsAg-positive persons in medical ex-amination who have no symptom. In addition, Pre-S1Ag in serum from 300 random samples of HBsAg-negative persons were detected in medical examination. Results Pre-S1Ag positive rate was 41.1% (351/854) in 854 samples with HBsAg(+) ,and HBeAg positive rate was 26.6% (227/854) in them. Pre-S1 Ag positive rate was 79.7% (181/227) in HBeAg(+) group but was 27.1% (170/627) in HBeAg(-) group. Pre-S1 Ag wasn't detected in 300 samples with HBsAg(-). Conclusions In HBsAg-positive samples who have no symptom,the detection rate of Pre-SIAg is higher than that of HBeAg. Pre-Sl Ag can reflect duplication and infectivity of HBV better than HBeAg and it has more value to epidemiology investigation and clinical diagnosis than the latter.
ABSTRACT
Objective To analyze and compare the Pre-S1 antigen and the serum markers to discuss the clinical significance of the Pre-S1 antigen of HBV.Methods The serum markers and the Pre-S1 antigen of HBV were detected by ELISA.Result By analyzing the serum markers in different combination modes,we found that the detection rate of the Pre-S1 antigen in different combination modes are:94% ( in the HBeAg positive mode) ; 15.6%(in the HBeAg negative mode) and negative in the mode both HBsAg and HBeAg are negative.Conclusions The Pre-S1 antigen of HBV has a coherence with HBeAg to a large extent,and has contact relations with the exist and replicate of HBV and can reflect the replication of HBV.The detection of the Pre-S1 antigen can help the diagnosis of HBV infection.
ABSTRACT
Objective To study the exact function of human gene 5 transactivated by pre-S1 protein of hepatitis B virus(PS1TP5)by investigating the gene expression of PS1TP5 in yeast cells. Methods Reverse transcription-polymerase chain reaction(RT-PCR)was performed to amplify the gene of PS1TP5 using the mRNA of HepG2 cells as template and the gene was cloned into pGEM-T vector.The gene of PS1TP5 was cut from pGEM-T-PS1TP5 vector and cloned into yeast expressive plasmid pGBKT7,then pGBKT7-PS1TP5 was transformed into yeast cell AH109.The yeast protein was isolated and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and Western hybridization.Results PS1TP5 gene was successfully amplified and identified by DNA sequencing.The digested fragment was cloned into pGBKT7 vector and transformed into yeast cell AH109.The results of SDS-PAGE and Western assay showed that the relative molecular weight of the expressed product was about 36 950,and PS1TP5 protein existed in yeast cells.Conclusion The findings suggest that PS1TP5 can be successfully expressed in yeast cell.
ABSTRACT
Objective To observe the clinical significance and laboratory diagnostic values of Pre-S1 antigens (Pre-S1)and its relativity to HBeAg and HBV-DNA after using interferon in association with kurarinone.Methods The content of Pre-S1 and HBV-M was detected by ELISA and the levels of HBV-DNA were detected by flores- cence quantitative PCR(FQ-PCR)in 100 serum samples of patients with HBV infection and patients after an antivi- ral treatment.Results Matched control in 50 serum of HBsAg-positive and detectable HBV-DNA cases,both 29 samples in 30 HBeAg-positive cases and 17 samples in 20 HBeAg-negative cases Pre-S1s were positive.Treatment set 50 serum of patients after taking two courses of an antiviral treatment for HBV infection,drawing blood to observe related markers,5 cases of effect set were turned into negative for serologic markers and virologic markers.19 cases of valid set,serologic markers were various and the HBV-DNA copies descended 2-3 orders of magnitude.26 cases of invalid set,serologic markers were not various,and 30 % samples of the HBV-DNA copies descended 1～2 orders of magnitude.Conclusion It was supplementary for Pre-S1 to indicate HBV expression when the HBeAg varied.It also provided detcctable laboratory markers in HBV infectivity,replication and therapeutic efficiency evaluation.
ABSTRACT
Objective To investigate the correlation among HBV infection models,HBV Pre-S1 antigen and HBV DNA loads in serum and to provide evidence for the diagnosis and treatment of hepatitis B.Methods Hepatitis B markers,HBV Pre-S1 antigen and HBV DNA loads in serum samples were detected simultaneously for 1037 cases.Results The positive rate of HBV Pre-S1 antigen was higher in HBsAg,HBeAg and HBcAb positive model than that in other models(P0.05).Conclusion The quantitative detection of HBV DNA is important for the prevention of HBV infection.The combination assay of serum HBV markers,Pre-S1 antigen and HBV DNA load was beneficial to the monitoring of disease condition of hepatitis B and to the improvement of therapeutic effect.
ABSTRACT
BACKGROUND: Ground glass hepatocytes are unique histological feature of chronic hepatitis B viral infection. The pre-S1 region of large surface protein has been shown to regulate assembly, processing, and secretion of HBsAg. The purpose of this study was to elucidate that a mutant form of pre-S1 affects this normal secretory pathway and is responsible for ground glass hepatocyte. METHODS: We examined HBV sequences spanning the pre-S region from a patients with HBeAg positive chronic HBV infection. HBV DNA was extracted from serum, cloned, and sequenced and determined the intrahepatic viral composition by extracting HBV DNA from paraffin embedded liver tissue. To analyze the viral population of single groundglass hepatocytes, we used the technique of laser capture microdissection to isolate individual hepatocytes from biopsy specimen. Groundglass hepatocytes that stained positively with anti-HBs and normal hepatocytes were harvested individually and their subjected HBV DNA sequences were analyzed. To define the responsible mutations for the HBsAg secretion, we introduced the mutant gene into molecular clone of wildtype (adwR9) and assayed their HBsAg amounts in the transfected cell supernatants by ELISA. RESULTS: Of 12 clones in serum analyzed, 9 clones had identical wild type sequences in the N-terminal region of the pre-S1 protein which plays an important role in the secretion and retention of HBV envelope proteins. One of the wild type clones has deletion within pre-S2 region. 3 identical mutant clones were isolated. Mutant type clones were predominant groundglass hepatocytes. CONCLUSIONS: We speculate that a mutant form of the HBV pre-S1 protein may result in the formation of ground-glass hepatocytes. Expression of abnormal pre-S1 may lead to its retention and accumulation within hepatocytes.
Subject(s)
Humans , Base Sequence , Biopsy , Clone Cells , DNA , Enzyme-Linked Immunosorbent Assay , Glass , Hepatitis B e Antigens , Hepatitis B Surface Antigens , Hepatitis B, Chronic , Hepatocytes , Laser Capture Microdissection , Liver , Paraffin , Secretory PathwayABSTRACT
Objective To evaluate the influence of expression of pre S1 protein on the genomic expression of hepatitis B virus (HBV) infected hepatocyte with microarray. Methods The differentially expressed genes between the hepatoblastoma cell line HepG2 transfected by pcDNA3.1(-) and pcDNA3.1(-)-preS1, were respectively compared by cDNA microarray technique. The HBV pre-S1 coding DNA fragment was amplified with polymerase chain reaction (PCR) technique by using G376-7 plasmid DNA containing the full length of HBV genome as the template. The expressive vector of pcDNA3.1-preS1 was constructed by routine molecular biological methods. HepG2 cells were transfected by pcDNA3.1(-) and pcDNA3.1-preS1, respectively, using FuGENE6 Transfection Reagent. The total RNA was isolated and reversely transcribed. Results The cDNAs were subjected for microarray screening with 1152 cDNA probes. From the scanning results, it was found that 30 genes were up-regulated and 38 genes were down-regulated by pre-S1 protein of HBV. Conclusion The expression of pre-S1 protein affected the genomic expression spectrum of HBV infected hepatocyte(HepG2 cell line).
ABSTRACT
PURPOSE: Hepatitis B virus(HBV) with various mutations has been reported. The aims of this study were to investigate the frequency and manifestation of HBV pre-S/S mutations in children with chronic hepatitis B infection. METHODS: Sera from 17 children with chronic hepatitis B infection were analyzed by direct sequencing of polymerase chain reaction amplification of HBV DNA. Results: Seventeen cases of adr type were analyzed. The deletions in HBV pre-S region were observed in 3(17.6%) of 17 cases. Of 3 deleted cases, 2 had an in-phase deletion in the pre-S1 region spanning 18 bp. Another case had a 18 bp and 3 bp deletions in the pre-S1 region. Many point mutations in HBV pre-S region were detected in all cases and these mutations were observed more frequently in the pre-S2 region than the pre-S1 region. Six point mutations in the pre-S1 region were observed. Eight point mutations in pre-S2 region were observed. Point mutations in the S region were detected in 14(82.4%) of 17 cases. Among these, mutations of the "a" determinant were detected in 4(23.5%) of 17 cases. Mutations at codon 130 and at codon 146 were noted in 2 cases. Combined mutations at codon 124, 126, 146 and at 130, 131, 136, 146 were noted in the other 2 cases. Mutations except "a" determinant region included at codon 3, 29, 73, 120, 184, 214, 226, 227. CONCLUSION: These observations suggest that deletion and point mutations in HBV pre-S1, pre- S2 regions and point mutations in HBV S region are frequent in the children with chronic hepatitis B infection.
Subject(s)
Child , Humans , Codon , DNA , Hepatitis B virus , Hepatitis B , Hepatitis B, Chronic , Hepatitis , Hepatitis, Chronic , Point Mutation , Polymerase Chain ReactionABSTRACT
To study the relationship between serum Pre-S1 antigen and HBV DNA, serum Pre-S1 antigen and HBV DNA marker in 229 patients positive for HbsAg were detected by using ELISA method and compared. It was found that both Pre-S1 antigen and HbeAg were correlated with HBV DNA with the coefficient being 0. 9812 and could be used to reflect the existence or reproduction of HBV DNA well. Although there was statistical correlation between them, their clinical implication was not completely the same. Therefore, a conclusion was drawn that combined detection of Pre-S1 antigen, HBV and HBV DNA in the patients with different kinds of HBV infection was helpful to clinical diagnosis, therapeutic effect evaluation and prognosis judgement.
ABSTRACT
Objective:To study the correlation between HBV Pre-S1 antigen,HBeAg levels and HBV DNA copies,so as to assess the clinical value of Pre-S1 in detection of HBV replication.Methods:A total of 363 HBsAg-positive samples were col- lected.The levels of Pre-S1 antigen,HBeAg and HBV DNA copies were determined by ELISA,time-resolved immuno-fluores- cent method and real-time fluorescent quantitative PCR(FQ-PCR),respectively.The correlation between the determination re- sults was analyzed.Results:Pre-S1 antigen level was correlated with the level of HBeAg(X~2=94.4,P
ABSTRACT
Objective To clone a new human gene 5 trans-activated by pre-S1 protein of hepatitis B virus (HBV), PS1TP5, and explore its structure and function by bioinformatics analysis. Methods PS1TP5 was amplified by reverse transcription-polymerase chain reaction (RT-PCR) technique by using HepG2 cDNA as template and inserted into pGEM-T vector by TA cloning. Recombinant eukaryotic expression vector pcDNA TM 3.1/myc-His A-PS1TP5 had been constructed by subcloning, followed by restriction enzyme digestion analysis and sequencing. Bioinformatic methods were used to analyze its possible physical and chemical characters, structure, and function. Results PS1TP5 was successfully amplified and cloned into pGEM-T and pcDNA TM 3.1/myc-His A vector by RT-PCR from HepG2 cDNA. The new gene had been confirmed by sequencing after PCR identification and restriction enzyme digestion and named as PS1TP5 because of its trans-active function. The sequence for the PS1TP5 gene had been deposited into GenBank, the accession number was AY427953. Bioinformatics analysis showed that its ORF was 438bp and translated a protein of 145 aa. Conclusion A new gene-PS1TP5 has been recognized, and its recombinant eukaryotic expression vector (pcDNA TM 3.1/myc-His A-PS1TP5) has been constructed. These results will certainly bring some new clues for the study of the biological function of new gene and pathogenesis of chronic hepatitis B.