Evaluation of ARCHITECT HCV Core Antigen Assay / 대한진단검사의학회지

BACKGROUND: Hepatitis C virus (HCV) core antigen (Ag) levels are known to be well correlating with HCV RNA levels, and may be used as an alternative marker of HCV replication for monitoring the response to HCV treatment. However, the low sensitivity of HCV core Ag assay has been an obstacle for clinical use. In this study, recently developed ARCHITECT HCV Ag assay (Abbott Laboratories, USA) was evaluated for analytical performance and clinical usefulness. METHODS: A total of 109 sera from HCV infected patients including various genotypes of HCV (1b, 2, 2a/2c, 2b, and 3a) and 20 sera from healthy donors were used for evaluating the sensitivity, precision, and linearity of the HCV core Ag assay. The cross reactivity with HIV, hepatitis B virus and myeloma proteins (N=5, each) and correlation with HCV RNA PCR assay were also evaluated. RESULTS: The sensitivity of the HCV core Ag assay was 97.2% (106/109) and there were no false positive results and cross reactivity. The within-run, between-run and between-day CVs were 3.0%, 2.5% and 3.0%, respectively. The levels of HCV core antigen showed a good correlation with those of HCV RNA quantification (r=0.940). The HCV Ag assay showed an excellent linearity in the range from 0.63 to 17,114 fmol/L (r=0.999). CONCLUSIONS: The ARCHITECT HCV Ag assay was good in sensitivity, precision, and linearity and its results well correlated with HCV RNA levels. This assay could be used as a good marker of viral replication for monitoring the therapy response in chronically HCV infected patients.

Production in Pichia pastoris and characterization of genetic engineered chimeric HBV/HEV virus-like particles / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the presentation of a neutralization epitope-containing peptide antigen of hepatitis E virus (HEV) on chimeric virus-like particles (VLPs) of hepatitis B surface antigen (HBsAg).</p><p><b>METHODS</b>The gene fragment corresponding to amino acids (aa) 551-607 (HEnAg) of HEV capsid protein, which contains the only neutralization epitope identified to date, was fused via a synthetic glycine linker in frame with the gene of HBsAg. The resulted fusion gene was then integrated through transformation into the genome of Pichia pastoris under the control of a methanol-induced alcohol oxidase 1 (AOX1) promoter and expressed intracellularly. The expression products in the soluble cell extracts were characterized by Western blot, ELISA, CsCl density gradient analysis, and electron microscopic visualization.</p><p><b>RESULTS</b>The novel fusion protein incorporating HBsAg and the neutralization epitope-containing HEnAg was expressed successfully in Pichia pastoris with an expected molecular weight of approximately 32 kD. It was found to possess the ability to assemble into chimeric HBV/HEV VLPs with immunological physical and morphological characteristics akin to HBsAg particles. Not only did the chimeric VLPs show high activity levels in a HBsAg particle-specific ELISA but they were also strongly immunoreactive with hepatitis E (HE) positive human serum in a HEV specific ELISA, indicating that HEnAg peptide fragments were exposed on VLP surfaces and would be expected to be readily accessible by cells and molecules of the immune system. Similarity between chimeric VLPs to highly immunogenic HBsAg particles may confer good immunogenicity on surface-displayed HEnAg.</p><p><b>CONCLUSION</b>The chimeric HBV/HEV VLPs produced in this study may have potential to be a recombinant HBV/HEV bivalent vaccine candidate.</p>

Particulate recombinant hepatitis E virus capsid protein and its antigenicity and immunogenicity / 生物工程学报

An E. coli expressed recombinant antigen NE2 was reported to aggregate into homo-oligomer, and can induce protective antibodies on rhesus monkey, but its immunogenicty was much weak after being purified. In this study, three N-terminal extension mutant of NE2 were expressed in E. coli, one of which named HEV 239 was found to aggregate into particle. HEV 239 antigen had good reactivity with sera of hepatitis E patients. The reactivity of HEV 239 against neutralization monoclonal antibody 8C11 was similar as NE2 antigen, while the reactivity of it against another neutralization monoclonal antibody 8H3 is much better than NE2 antigen, which indicated better antigenicity of HEV 239 than NE2. The diameter of purified HEV 239 particulate antigen was between 15 nm to 30 nm. The ED50 of immunization of HEV 239 particle adsorbed by aluminum adjuvant to BALB/c mice was between 0.08 microg to 0.25 microg. In contrast, the seraconversion rate of mice immunized by NE2 antigen adsorbed by aluminium adjuvant was only 25% on 60 microg vaccination. These results suggested that HEV 239 antigen particle has better immunogenicity as well as antigenicity than those of NE2 antigen, so it is a better vaccine candidate against HEV.

A serosurvey of hepatitis B virus, hepatitis C virus, human T lymphotropic virus type-1 and syphilis in HIV-1-infected patients in Jamaica

The seroprevalences of hepatitis B virus (HBV), hepatitis C virus (HCV), human T lymphotropic virus type-1 (HTLV-1) and syphyilis were determined in 129 HIV-1-infected patients using commercially prepared reagents. The seroprevalences were HCV, 0 (0/129); HBV, 37 (48/129); HTLV-1, 5 (6/129) and syphilis, 20 (26/129). Fifteen per cent (19/129) of the patients had active/chronic HBV infection. The seroprevalence of HBV was statistically significantly higher in HIV-1 infected men (24/49, 50 versus 17/80, 21; p = 0.005), while the seroprevalence of syphilis was statistically significantly increased in HIV-1 infected patients in the over-40 age group (10/31, 32 versus 6/53, 11; p = 0.05). These findings throw the spotlight on HBV infection and syphilis and suggest that these two sexually transmitted infections should be carefully surveyed in patients with HIV/AIDS in Jamaica. It is essential for management protocols in Jamaica to include screening for evidence of these co-infections

Construction of a recombinant human adenovirus expressing the ORF2 antigen of HEV and immunization of mice by mucosal system / 中国医学科学院学报

<p><b>OBJECTIVE</b>To construct a replication-defective recombinant adenovirus expressing the ORF2 (112-660aa) antigen of hepatitis E virus (HEV) and evaluate its immunization effect in BALB/c mice by mucosal inoculation.</p><p><b>METHODS</b>The HEV ORF2 gene encoding for 112-660aa was amplified from plasmid pUC-HEV and inserted into the transfer vector pTrack-CMV. The recombinant plasmid and adenoviral backbone plasmid pAdEasy-1 were co-transformed into E. coli strain BJ5183. Taking the advantage of the high efficient homologous recombination machinery presented in bacteria, the recombinant adenovirus backbone plasmid was generated in BJ5183, and then was transfected into 293 cells. Recombinant Adenoviruses were propagated in 293 cells with high titers. 8-week-old BALB/c mice were inoculated intraperitoneally and intranasally with 10(7) pfu recombinant adenovirus each on weeks 0, 3, 5, 7, 10.</p><p><b>RESULTS</b>Both groups of mice induced humoral IgG immune response with the highest titers 1:1,000 and 1:10,000 each. Only the group inoculated intranasally could induce mucosal IgA immune response.</p><p><b>CONCLUSIONS</b>The adenoviral recombinant can stimulate specific humoral and mucosal immune response in mice and is potentially to be used as a candidate vaccine for the treatment of HEV infection.</p>

Antigencity identification of recombinant hepatitis E virus ORF2 protein expressed in Pichia pastoris / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To determine the antigenicity of recombinant hepatitis E virus ORF2 (rHEV ORF2) protein expressed in Pichia pastoris (P. pastoris).</p><p><b>METHODS</b>By using the rHEV ORF2 protein from E.coli as control, an indirect ELISA was adopted to identify the sensitivity, specificity and stability of rHEV ORF2 protein from P. pastoris in detection of HEV IgM and IgG antibody in sera from patients with hepatitis E. The reactivity of the rHEV ORF2 against 5 HEV ORF2 monoclonal antibodies (McAbs) was also tested.</p><p><b>RESULTS</b>The minimum concentration of coated antigen with which HEV IgG could be detected was 12.5 ng/ml, while the highest serum dilution to detect both IgM and IgG antibodies against HEV was 1:5 120. No cross-reaction was found with sera from patients with any other types of hepatitis. The 37 degree C acceleration test showed that the rORF2 was highly stable within 12 months at 4 degrees C. The 5 HEV ORF2 McAbs showed better reaction with the rORF2 from P. pastoris, especially that 4B2, 2E2, whose reaction against the rORF2 were 125 and 25 times respectively higher than that of rORF2 from E.Coli.</p><p><b>CONCLUSION</b>There may be more extensive conformational epitopes in the rHEV ORF2 from P. pastoris. The excellent antigenicity, sensitivity and stability suggest that it can be served as a new candidate antigen for the development of diagnostic reagents of hepatitis E.</p>

Role of diet in management of different forms of chronic hepatitis in young adolescents

The role of diet in treatment of chronic hepatitis in young adolescents was explored. About 110 patients with chronic hepatitis were classified into three groups: Group I included 46 patients with chronic hepatitis C, group II included 37 patients with chronic hepatitis B and group III included 27patients with chronic hepatitis due to other causes rather than C or B infection. Liver biopsy was done for all patients. Anthropometric parameters and blood samples were taken before and after diet program for six months. The biochemical parameters revealed a significant increase in the serum level of total proteins and hemoglobin concentration, while there was a significant decrease in total and direct bilirubin, alkaline phosphatase, ALT and AST. Non significant differences were demonstrated in body weight, height, arm and wrist circumferences, WHR and BMI in patient groups after diet program

Relationship between amount of HBV DNA in serum/liver tissue and hepatitis G virus (HGV) infection in patients with chronic hepatitis B / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To observe the relationship between the amount of HBV DNA in serum/liver tissue and HGV infection in patients with chronic hepatitis B (CH-B) for exploring the effect of HGV infection on hepatitis B virus (HBV) replication of CH-B.</p><p><b>METHODS</b>HGV RNA in serum, HGV nonstructural region 5 (NS5) antigen (HGV Ag) in liver tissue and the amount of HBV DNA in serum, liver tissue were detected for 56 patients with CH-B by reverse transcription-polymerase chain reaction (RT-PCR) assay, peroxidase antiperoxidase (PAP) immunohistochemical method and fluorescence quantitative PCR assay, respectively. Then the relationship between HGV Ag expression in liver tissue and HGV RNA expression in serum was analysed and the amount of HBV DNA in serum and liver tissues from the serum HGV RNA or liver tissue HGV Ag positive patients were compared with those of the serum HGV-RNA or liver tissue HGV Ag negative patients, respectively.</p><p><b>RESULTS</b>Ten (17.9%) and eight (14.3%) patients were positive for serum and liver tissues,respectively.HGV RNA expression in serum was closely related to HGV Ag expression in liver tissues, but there was HGV RNA in serum from some of the liver tissues HGV Ag negative patients ?cases of HGV RNA and HGV Ag positive or negative,HGV RNA positive but HGV Ag negative, HGV RNA negative but HGV Ag positive, respectively: 5,43,5,3,(P<0.01). There was no significant difference in the amount of HBV DNA in serum and liver tissues between HGV RNA or HGV Ag positive and negative patients (P>0.05).</p><p><b>CONCLUSIONS</b>HGV infection may not affect HBV replication. Liver is the site of HGV replication, but HGV probably also replicates in extrahepatic tissues. HGV hepatic pathogenicity is probably mild and further studies are still needed.</p>

Histological comparison of chronic hepatitis B and C in an Indian population.

Many studies have shown that steatosis, lymphoid aggregates or follicles and bile duct injury on histology are more consistently associated with chronic hepatitis C than chronic hepatitis B. We compared liver biopsies of 30 patients of chronic hepatitis B with an equal number of age matched patients with chronic hepatitis C. Steatosis, lymphoid aggregates or follicles and bile duct injury were noted in 66.6%, 36.6% and 26.6% cases respectively of chronic hepatitis B as compared to 70%, 33.3% and 30% cases respectively of chronic hepatitis C. Thus none of the features were considered distinctive of HCV infection.

Incidence of hepatitis A, B, and C in Embaba fever hospital among acute hepatitis patients

This work was conducted on 107 adult Egyptian patients with acute viral hepatitis as well as ten normal age and sex matched subjects taken as a control group. The patients were divided into HAV, HBV, HCV, combined HBV and HCV and seronegative groups. The results indicated that among 107 patients diagnosed as acute viral hepatitis, 55 patients were positive for HbsAg, 28 patients were positive for HCV antibodies and seven patients were positive for HAV. Also, six patients were positive for both HBV and HCV and 11 patients were seronegative. The incidence of bilharziasis was higher in the seronegative group. It was concluded that the most frequent cause of acute viral hepatitis among the studied Egyptian patients was HBV, followed by HCV infection; while HAV represented the least one to cause acute viral hepatitis among adults

Antigen profile among patients with autoimmune hepatitis

Autoimmune hepatitis [AIH] is a disease of an unknown cause. Our study was conducted on 20 patients diagnosed by laboratory according to Mackay's criteria 15 as having Autoimmune hepatitis and 20 healthy controls. Each patient and control were fully typed for HLADRB class II alleles using sequence specific probes [SSP] polymerase chain reaction [PCR] typing. Our results detected that DR[4] alleles was highly significant detected among patient [48.8%] in contrast with [6.3%] among controls [RR, 7.8 and P<0.001]. Also the subtype 0401 was the highly prevalent subtype detected among patients and controls [50% and 3.8%], respectively [RR 13.5 and P<0.001]. Also DR[15] allele was negatively associated with AIH [7.5% and 37.5%] among both patients and controls, respectively. It is considered as a protective alleles. Furthermore, the subtype 1501 was highly prevalent among controls [48.8%] in contrast [6.3%] among patients. Most patients were females with hypergammaglobulinemia and characteristic autoantibodies were diagnostic markers. In conclusion DR[4] allele is the most prevalent associated allele in AIH patient and the subtype 0401 is a prevalent significant subtype. Furthermore, DR[15] is the most prevalent protective allele and its subtype 1501 is the most predominant subtype. HLA typing is an easy and useful method for detecting AIH patients so early management with immuno supervise therapy in order to prevent the liver damage

Hepatitis virales / Viral hepatitis

En esta revisión se describen los virus hepatotropos actualmente conocidos, su epidemiología con referencia especial a los datos nacionales relativos a los virus A, B, C y E; su historia natural y sus aspectos clínicos más relevantes. Se enfatizan, además los diferentes marcadores virales serológicos, para el diagnóstico de infección aguda o crónica. Se incluyen también los diferentes tratamientos y las medidas de prevención (pasivas o activas) recomendadas actualmente

Hepatites virais: atualizaçäo / Viral hepatitis: update

Objetivo: os autores apresentaram uma revisäo dos diferentes tipos de hepatite viral na infância. Säo discutidos, com ênfase especial, os aspectos clínicos-laboratoriais e evolutivos das formas mais freqüentes da doença. Métodos: Foram selecionados, através de Medline, os artigos mais significativos publicados nos últimos anos sobre hepatite viral na infância. As características epidemiológicas säo discutidas, sempre que possível, levando em conta dados brasileiros. Resultados: A presente revisäo analisa as hepatites causadas por agentes que têm em comum a origem viral e o hepatotropismo mas que determinam doenças com diferenças marcantes sobretudo quanto a evoluçäo e prognóstico. Em relaçäo ao vírus B, por exemplo o paciente pode apresentar desde um simples estado de portador até hepatite aguda, hepatite crônica, cirrose...

Prevalência dos marcadores sorológicos dos vírus da hepatite B e D em crianças das tribos Caiabi e Txucarramäe do Parque Indígena do Xingu, Brasil central / Prevalence of serological markers of hepatitis B and D viruses in Indian children of the Caiabi and Txucarramäe tribes of Xingu Indian Park in Central Brazil

The medical literature has shown that the inhabitants of the Amazon region are highly affected by hepatitis B and delta viruses infection, but this has never been studied in Indian children by age group. A study of the prevalence of serological markers of hepatitis B and delta viruses in Indian children aged 0 to 14 years living in the PIX was carried out. This Park is located in Central Brazil, a region which is in the transition between the savannab to the south and the Amazon jungle to the north. To determine the prevalence of HBV and HDV markers in Indian children and to characterize the route of HBV transmission in this region. Out of the 17 tribes living in the PIX, two--the Caiabi and the Txucarramäe--were chosen because both live in the North part of the Park, but have quite different ways of life. The overall prevalence of HBV serum markers was: HBsAg, 4.5 per cent; anti-HBs, 39.6 per cent; anti-HBc, 44.1 per cent; any marker of HBV, 47.3 per cent; and anti-HDV, 0.0 per cent. However, a striking difference in the prevalence of hepatitis B markers was observed between the two tribes: younger Caiabi children were much less affected than the Txucarramäe ones. The prevalence of HBsAg in fertile women was 12 per cent, being anti-HBe positive. Our data suggest that HBV infection is highly prevalent among Indian children living in this Indigenous Park and vertical infection is not an important route of transmission in either tribes.

Epidemiology of hepatitis delta virus infection in HBs Ag positive patients

Over a period of three years in Medical Unit I Civil Hospital Karachi, 230 patients screened positive for HBsAg, out of these 52 patients showed up anit-Delta antibodies giving a prevalence of 22.6 percent. 45/52 i.e. 86.53 percent were male and 7/52 [13.47 percent] were female thus indicating a male predominance. All age groups are affected by HDV infection, but prevalence was highest in patients in the sixth decade followed by the third decade. Meticulous history taking showed the probable routes of transmission. 16/52 [30.7 percent] revealed a history of parenteral injections, 10/52 [19.32 percent] had undergone a blood transfusion, 6/52 [11.53 percent] were drug addicts; 6/52 [11.53 percent] revealed sexual promiscuity, and 2/52 [3.82] patients underwent hamodialysis. In addition, majority of patients with evidence of HDV infection came from rural Sindh or far flung areas of Baluchistan. In conclusion, prevalence of HDV infection was found to be higher than that reported earlier from western countries and equal to or even higher than other Afro-Asian countries. The association of infection with probable routes of transmission is very strong in our patients. Patients from rural areas are more prone to develop this sinister infection

Efectividad terapéutica de la S-Adenosil-L-Metionina en hepatitis viral aguda A,B, NANB

La hepatitis viral aguda es una causa frecuente de enfermedad en el mundo, ocasionado serias complicaciones, y altos costos como consecuencia de la incapacidad resultante. A pesar de esto, no disponemos de ninguna modalidad terapéutica de probada eficacia. El papel exacto de la SAMe en el tratamiento de las hepatopatías ha sido parcialmente clarificado por estudios recientes que muestran, que esta molécula es de efecto benéfico en la colestasis intrahepática (DRUGS 0:111, 1990) y otras hepatopatías. Para evaluar la efectividad de la SAMe se realizó un estudio doble ciego, placebo, control en 28 pacientes entre enero 25 de 1989 y junio 15 de 1991 con los siguientes criterios de inclusión: Edad entre 10 y 65 años, evidencia clínica de hepatitis aguda A, B o NANB, menos de 15 días de evolución, AST y-o ALT mayor de 500 U-L y bilirrubina total mayor de 3 mg-dl. Todos los pacientes fueron aleatorizados para recibir SAMe 500 mgr IV diariamente por 10 días o Placebo (l-Lisina 150 mg; Hidróxido de sodio 4.5 mgs; agua destilada estéril 5.0 ml) IV diariamente por 10 días. Ambos grupos eran comparables en relación con la edad, etiología y pruebas hepáticas. Las diferencias entre los grupos se evaluaron utilizando las pruebas ANOVA, t de Student, con programa estadístico SPSS-PC. Un valor de menos de 0.005 se consideró significativo