Purification effect of composite chromatography medium Capto Core 400 on hepatitis A virus / 中国生物制品学杂志

@#ObjectiveTo explore the purification effect of hepatitis A virus(HAV)with a new type of composite medium Capto Core 400 chromatography packing.MethodsHAV crude liquid cultured with human embryonic lung diploid cells(2BS strain)was purified using Capto Core 400 chromatography media. The optimum conditions for purification of HAV were determined from flow rates(60,150,300 and 450 cm/h),pH of loading buffer(6. 6,7. 0 and 7. 4),salt ion concentration of loading buffer(0,0. 15,0. 3 and 0. 45 mol/L)and capacity[0. 5,1,2 and 4 column volume(CV)],respectively. The purification effect was compared with that of traditional Sepharose 4FF chromatography media.Results The optimum purification conditions of Capto Core 400 were preliminarily determined as follows:flow rate of 150 cm/h,loading buffer pH of 7. 0 ～ 7. 4,salt ion concentration of 0. 15 mol/L,loading volume of 2 CV. The average recovery rate of virus was 92. 61%,and the average removal rate of protein was 84. 25%. The average recovery rate of HAV antigen purified by Sepharose 4FF chromatography media was 76. 03% and the average protein removal rate was 73. 21%. The purification effect of Capto Core 400 chromatography media was better than that of Sepharose 4FF chromatography media.ConclusionThe Capto Core 400 composite chromatography media had a high recovery rate for HAV purification and a relatively simple operation,which was suitable for HAV purification.

Prokaryotic expression and immunogenicity of hepatitis A virus capsid proteins VP1 and VP3 / 中国生物制品学杂志

@#Objective To express the capsid proteins VP1 and VP3 of hepatitis A virus(HAV) in prokaryotic cells and evaluate their immunogenicity.Methods VP1 and VP3 gene fragments were amplified by PCR,cloned into vector pETG28a to construct recombinant expression plasmids pET-G28a-VP1 and pET-G28a-VP3,which were transformed into competent E.coli BL21(DE3),induced by IPTG,and then analyzed by 12% SDS-PAGE.The target protein was purified by ion exchange chromatography,renatured and combined with several adjuvants to immunize mice.The mice were divided into VP1-MF59,VP1-AH,VP1-AP,VP1-AP-10CpG,VP1-AP-50CpG,VP3-AP,VP3-VP1-AP and PBS control groups,five for each group.Serum IgG antibody titers of mice in various groups were detected by ELISA,and serum neutralizing antibody titers of mice in VP1-AP,VP3-VP1-AP and PBS control groups were detected by rapid fluorescence focus immunosuppression experiment.Results Colony PCR and sequencing showed that the recombinant plasmids pET-G28a-VP1 and pET-G28a-VP3were constructed correctly.The recombinant proteins VP1 and VP3,with relative molecular masses of about 37 000 and26 000 respectively,mainly existed in the form of inclusion bodies,the expression levels were 18.6% and 32.4%,and the purity was 86.3% and 84.7%,respectively.The recombinant proteins VP1 and VP3 reacted specifically with rabbit anti-HAV antiserum and mouse anti-HAV-VP3 antiserum respectively.The serum IgG antibody titer of mice in VP1-AP-50CpG group was significantly higher than that in VP1-AP group(q=22.05,P <0.01),and the serum IgG antibody titer of mice in VP3-VP1-AP group was significantly higher than that in VP1-AP group and VP3-AP group(q=22.05 and 22.49 respectively,each P <0.01).Compared with PBS control group,the serum neutralizing antibody titer of mice in VP1-AP and VP3-VP1-AP group increased significantly(q=7.79 and 25.11 respectively,P<0.01) Conclusion Prokaryotic HAV capsid proteins VP1 and VP3 showed high purity and good immunogenicity,and the addition of CpG in the preparation was beneficial to enhance the immunogenicity of antigens.This study laid a foundation of the development of HAV recombinant subunit vaccine.

Optimization for condition of adsorbing hepatitis A virus antigen with aluminum hydroxide adjuvant and evaluation of immunogenicity of adsorption products / 中国生物制品学杂志

@#Objective To optimize the adsorption condition of hepatitis A virus(HAV)antigen by aluminum hydroxide[Al(OH)_3]adjuvant and evaluate the immunogenicity of the adsorbed products. Methods The final molar concentration of buffer(0. 01,0. 02 and 0. 04 mol/L),pH value(5. 5,6. 0,6. 5,7. 0,7. 5 and 8. 0),final molar concentration of sodium chloride(NaCl)(0. 075,0. 13,0. 15,0. 17,0. 45,0. 75 and 1. 0 mol/L),mixing mode(mixing after dilution and mixing directly without dilution)and dropping order[HAV stock solution into Al(OH)_3 solution,and Al(OH)_3 solution into HAV stock solution]were selected for single factor screening. Taking Al(OH)_3 adsorption rate as the evaluation index,the selected factors were optimized by orthogonal test. NIH mice were immunized intraperitoneally with adsorbed products prepared under optimal and suboptimal combination conditions with 0. 5 mL/mouse,10 mice in each group with half male and half female. After 28 d of immunization,the blood samples were collected from eyeball or heart,and the serum was separated. The HAV antibody level was detected by ELISA,and the immunogenicity of adsorbed products was evaluated by the seroconversion rate of mouse serum antibody. Results The results of single factor screening test showed that the final molar concentration of buffer,pH value and final molar concentration of NaCl had significant influence on the adsorption effect(F = 6. 582,14. 007 and 8. 612,respectively,each P < 0. 05),while the mixing mode and dropping order had no effect(t =-0. 696 and 1. 153,respectively,each P > 0. 05). The final molar concentration of buffer was 0. 01 and 0. 02 mol/L,the pH value was 5. 5 ～ 7. 0,and the final molar concentration of NaCl was 0. 13 ～ 0. 17 mol/L. The orthogonal test showed that the order of the influence on adsorption effect was the final molar concentration of buffer > pH value > NaCl final molar concentration. The optimal combination was pH 6. 3 + final molar concentration of buffer 0. 01 mol/L + NaCl molar final concentration 0. 15 mol/L,and the suboptimal combination was pH 5. 8 + final molar concentration of buffer 0. 02 mol/L +NaCl final molar concentration 0. 17 mol/L. The seroconversion rate of serum antibody in mice immunized with the adsorbed products prepared by the optimal and suboptimal combination was both 100%. Conclusion The adsorbed products prepared under the optimized conditions have good immunogenicity,which lays a foundation of the research of inactivated hepatitis A vaccine

Adaptability and genetic stability of hepatitis A virus SYX_1 strain in human diploid cell MRC-5 / 中国生物制品学杂志

@#Abstract：Objective To investigate the adaptability and genetic stability of hepatitis A virus（HAV）SYX1 strain in human diploid cell MRC⁃5. Methods HAV SYX1 strain isolated from feces of patients with hepatitis A was continuously propagated in MRC⁃5 cells for 28 passages，of which the 1st ~ 26th passages were determined for antigen contents and virus titers，the 6th passage was observed for the morphology under microscope and detected for physicochemical properties，and the 13th ~ 15th passages were studied for virus proliferation dynamics to determine the peak yield of virus proliferation. Genomic RNA was extracted from the 8th，12th，18th，20th，22nd，25th，26th and 28th passages and sequenced to analyze the genetic stability. The main seed batch and working seed batch of HAV SYX1 strain were established and verified according to the requirements of Chinese Pharmacopoeia（VolumeⅢ，2020 edition）. Results The antigen content of HAV SYX1 was stable at 160 ~ 320 EU／mL and the titer was maintained at 7. 3 ~ 8. 3 lgCCID50／mL after the 8th passages in MRC 5 cells；Virus particles showed two types：hollow and solid，with a diameter of 27 ~ 32 nm，spherical，without envelope and protrusions on the surface，which tolerated low pH value and ether. The peak period of virus proliferation was 10 d with an antigen content of more than 160 EU／mL and a virus titer of more than 7. 0 lgCCID50／mL. HAV SYX1 was a subtype of HAV IB，and no mutation in the coding region of all structural proteins during passage was observed. The verification results of main seed batch and working seed batch of HAV all met the relevant requirements. Conclusion HAV SYX1 strain showed good adapt⁃ ability and genetic stability in MRC⁃5，which might be used for the development and production of inactivated hepatitis A vaccine.

High prevalence of hepatitis A and E viruses in environmental and clinical samples from West Argentina

ABSTRACT Environmental surveillance of water sources is important to monitoring viral hepatitis transmission in clinical settings. This study investigated the circulation of hepatitis A (HAV) and E (HEV) viruses in sewage and clinical samples from Argentina. Between 2016 and 2017, 80 raw sewage samples and 86 clinical samples (stool and serum) from suspected cases of hepatitis A and hepatitis E were obtained. HAV and HEV were tested by both real-time and nested PCR. Positive samples were sequenced for genotype determination and phylogenetic analysis. Overall, HAV was recovered in 39% of sewage samples and 61.1% of clinical samples. HEV was detected in 22.5% of sewage samples and 15.9% of clinical samples. HAV was found more frequently in sewage during the winter and in clinical samples in spring; HEV was more prevalent in sewage during summer and in clinical samples in autumn. All HAV isolates belonged to genotype IA and HEV isolates belonged to genotype 3, the most prevalent genotypes in South America. High prevalence of HAV and HEV in environmental and clinical samples in Mendoza, Argentina was observed. These findings reinforce the importance of environmental surveillance and implementation of health strategies to control the spread of HAV and HEV in developing countries.

Demographic and Clinical Profile Analysis of Acute Viral Hepatitis

Background: Acute viral hepatitis (AVH) is a major health concern in developing countries like Bangladesh regarding morbidity as well as mortality. Usually, acute infections are caused by A, E, hepatitis viruses, and occasionally hepatitis B virus. Infection caused by the hepatitis C virus is usually asymptomatic. Prior knowledge of the demographic and clinical profile of acute viral hepatitis may be helpful for treatment professionals in the management of such patients. Aim of the study: The aim of this study was to assess the demographic and clinical profile of acute viral hepatitis patients in Bangladesh.Material & Methods:This prospective observational study was conducted in the Department of Gastroenterology, US-Bangla Medical College & Hospital during the period from March 2018 to February 2019, in Bangladesh. A total of 59 suspected patients with acute viral hepatitis were included as the study subjects for this study. Ethical approval of the study had been taken from the ethical committee of the mentioned hospital. A predesigned questionnaire was used in data collection. Collected data analyzed by using MS Office and SPSS version 23.0 programs as per need. A P-value, of <0.05 was considered significant.Results:Among 59 participants, the male-female ratio was 3.2:1 and the mean (±SD) age was 23.88±14.83 years. Analyzing hepatitis infection, we found hepatitis E was the highest in number 32(54.24%), followed by hepatitis E virus 20(33.9%), and hepatitis B virus 5(8.47%). In this study, we did not find any patients with hepatitis C virus, and 2 participants didn’t have any hepatotropic virus. The mean (±SD) Serum bilirubin (mg/dl) was 6.35±1.63, and the mean (±SD) serum alkaline phosphatase (IU/L), serum. creatinine (mg/dl), serum albumin (gm/dl) and plasma glucose random (mg/dl) were found 366.81±257.20, 6.97±14.96, 19.75±22.98 and 10.08±5.49 respectively. Considering dual viruses among the total of 5 patients along with hepatitis E & A viruses in this study, we found all were hepatitis B viruses. Among them, 3 with hepatitis E and the rest 2 were with hepatitis A virus consequently. Among them 3(60%) with hepatitis A and the rest 2(40%) with hepatitis E. In the issue of patients attended with clinical complaints, vomiting was the highest among hepatitis A patients 19(86.4%) followed by jaundice 8(36.4%) and fever8(36.4%). On the other hand, among hepatitis E patients’ jaundice was highest at 19(61.3%) followed by vomiting at 17(54.8%), and fever at 6(19.4%) respectively.Conclusions:The incidence of HEV is found as the most predominant among all the acute viral hepatitis patients and vomiting and jaundice were the most common presenting complaints.

Prevalence of hepatitis A virus and hepatitis E virus in the patients presenting with acute viral hepatitis in Rohtak, Haryana, India

Background: Hepatitis A virus (HAV) and hepatitis E virus (HEV) cause acute hepatitis in humans and are transmitted mainly through the fecal-oral route. They pose major health problems in developing countries. This study was done to determine prevalence of HAV and HEV in patients presenting with AVH and the co-infection of HAV and HEV in these patients.Methods: The study was conducted in the virology research and diagnostic laboratory, PGIMS Rohtak during the study period of August 2017-December 2018. The study population included sera of individuals from all age group who were suspected of acute viral hepatitis (AVH). All the sera were screened for IgM antibody to HEV and HAV using IgM capture ELISA.Results: HEV IgM ELISA test   was performed in 307 patients (mean age 34 years;), with an overall seroprevalence rate of 138(44.9%). HAV antibodies were detected in 109 subjects, with a median age of 9.5 years the seroprevalence of HAV was 34 (31.1%). HEV seropositivity was highest in the age group 20-30 years. Mean age was 34 years whereas the interquartile range was from 14-71 years. HAV infection was positive mainly in the age group <10 years. With interquartile range from 6-16 years. Out of total 34 patients positive for HAV infection males were 20 (58.8%), whereas females were 14(41.1%). HEV IgM was positive in 138 patients, out of which male were 96 (69.56%) and females were 42 (30.43%). HEV IgM was positive in 138 patients, out of which male were 96 (69.56%) and females were 42 (30.43%). HAV and HEV seen to be prevalent all with highest predominance seen towards the end of monsoons (August and September) and beginning of winters.Conclusions: The present study also points toward HEV being the prime etiological agent for outbreaks of acute hepatitis in the studied region of Haryana (Rohtak), India. A comparatively lower HAV prevalence may be the consequence of an overall declining trend due to improved living standards and environmental hygiene.

Characterization analysis and overexpressed vector construction of the circular RNA hsa_circ_0082626 derived from the antiviral gene ZC3HAV1 / 医学研究生学报

Objective Circular RNA is a research hotspot of non-coding RNAs. The purpose of this study was to investigate the basic characteristics and expression effect of the overexpression vectors of circular RNA hsa_circ_0082626 transcribed from antiviral gene ZC3HAV1. Methods The basic characteristics of hsa_circ_0082626 were studied by the verification of reverse cleavage site, RNase digestion assay and intracellular distribution location with extraction of cytoplasmic and nuclear RNA. In the RNase digestion assay, samples were divided into RNase R treatment group and control group. RNase. 20 U (2 U/μg) of RNase R was added to the R treatment group, and the control group was replaced with an equivalent amount of ddH2O to detect changes in expression levels after RNase treatment. The cells were divided into 2 groups: overexpression group and negative control group. At 24, 48 and 72 h after transfection, cells were collected to detect the expression of circular RNA by RT-qPCR. Results Compared with the control group, the expression of ZC3HAV1, CDR1as and GAPDH in the RNase R treatment group was increased (0.144±0.002 vs 1.000±0.016, 0.772±0.058 vs 1.000±0.122, 0.077±0.009 vs 1.000±0.164, P<0.05). Hsa_circ_0082626 could resist the treatment of RNase R and was mainly distributed in cytoplasm. The expression level of hsa_circ_0082626 in the overexpression group was significantly higher than that in the negative control group, and the expression level was the highest at 48 h after transfection. Conclusion The characteristics of hsa_circ_0082626 reverse cleavage site, RNase resistance and expression in cells were successfully analyzed, which proved that hsa_circ_0082626 does have a circular structure. The overexpression vector of hsa_circ_0082626 was successfully constructed to provide an experimental basis for the biological function and mechanism of RNA hsa_circ_0082626.

Initial evaluation of universal immunization with a single dose against hepatitis A virus in Central Brazil

ABSTRACT Vaccination against the hepatitis A virus (HAV) administered in two doses has been used effectively in universal child immunization programs in several countries. A single-dose vaccination was adopted in some low-income countries in an attempt to reduce costs without losing effectiveness. In 2014, single-dose universal vaccination was introduced in Brazil for children aged two years. Since such strategy is still not universally accepted, its efficacy should be compared to the two-dose strategy. To assess the humoral response after the single-dose HAV vaccination schedule, a cross-sectional study was conducted in Primavera do Leste, in Mato Grosso state, Central Brazil, including 265 children vaccinated through the National Immunization Program. Blood was collected by using a digital puncture and further applied to filter paper cards. Anti-HAV was detected in 218 out of 265 dried blood spots (DBS). Blood venous samples were collected from 34 out of 47 children who were not anti-HAV positive in DBS samples. Eighteen of them tested positive for anti-HAV, giving a final score of 93.6% (236/252) of seropositivity. In conclusion, this study demonstrated a high rate of anti-HAV positivity in the short term after single-dose hepatitis A vaccination in the population investigated. Moreover, the DBS was shown to be a reliable tool for detecting anti-HAV antibodies.

Molecular characterization of hepatitis A virus from children hospitalized at a tertiary care centre in northwest India

Background & objectives: Hepatitis A virus (HAV) infection is a major cause of childhood hepatitis, prevalent worldwide. HAV is classified into seven genotypes I-VII; genotypes III and I are the most common among humans. The present work was carried out to identify the genotypes prevalent in children suspected to have acute viral hepatitis (AVH), hospitalized at a tertiary care centre in northwest India. Methods: A total of 1269 blood samples from children (0-15 yr of age) clinically suspected of viral hepatitis were screened for anti-HAV IgM. Acute phase serum was processed for RNA extraction and amplified by nested polymerase chain reaction (PCR) followed by sequencing of representative samples. Results: Among the 1269 samples tested, 642 (50.59%) were positive for anti-HAV IgM; among the positive samples, 171 patients having a history of less than seven days were tested by PCR, of whom 141 (82.45%) were found to be PCR positive. Nucleotide sequencing of a representative 44 samples showed high homology; all the samples were found to be of genotype IIIA. Interpretation & conclusions: Hepatitis A was prevalent during July to September and in predominantly children less than five years age. Only genotype IIIA was detected in all the samples.

Detección del genotipo 1d del virus de la hepatitis E en pacientes con sospecha de hepatitis viral aguda, Cuba 2013 / Detection of hepatitis E virus genotype 1d in patients with suspect of acute viral hepatitis in Cuba during 2013

Resumen Introducción El virus de la hepatitis E (VHE) se transmite, principalmente, por vía fecal-oral y es una de las principales causas de hepatitis viral aguda (HVA) en el mundo. En Cuba, a pesar de que este virus tiene un comportamiento endémico, no se relaciona a este patógeno al presentarse una hepatitis viral de trasmisión entérica. Objetivo Teniendo en cuenta que el VHE y el virus de la hepatitis A (VHA) comparten rutas de transmisión, nos propusimos estimar la incidencia de ésta infección (VHE), en muestras que se recibieron de todo el país durante el año 2013, cuyo criterio de inclusión fue la indicación médica de IgM anti-VHA. Materiales y Métodos Se empleó la RT-PCR específica para el marco abierto de lectura 2 (MAL2), con el propósito de detectar el ARN-VHE en las 422 muestras estudiadas. Los productos amplificados fueron purificados, secuenciados y analizados filogenéticamente con el programa MEGA6. Resultados La presencia de ARN-VHE se detectó en 8,53% (36/422) de las muestras estudiadas. El mayor índice de positividad se identificó en la región occidental del país, específicamente en La Habana con 5,45% (23/422). En total se diagnosticaron 5,21% (22/422) muestras positivas al marcador de IgM VHA; la detección simultánea de marcadores del VHA-VHE fue 13,88% (5/36). Los resultados demuestran una mayor incidencia del VHE con respecto al VHA (8,53% vs 5,21%) y el análisis filogenético mostró la circulación del genotipo 1, subgenotipo 1d del VHE. Conclusiones Se corroboró la endemicidad del VHE en nuestro país y, por primera vez, se identificó el subgenotipo 1d, variante africana asociada a casos esporádicos y brotes de hepatitis E.

Abstract Introduction Hepatitis E virus (HEV) is mainly transmitted by the fecal-oral route and is one of the most important causes of acute viral hepatitis (AVH) around the world. In Cuba, Despite of endemic behavior of HEV in Cuba, its causality is not associated when a picture of enteric acute hepatitis is suspected. Objective Taking into account the common transmission route of both HEV and HAV, our aim was to estimate the incidence of HEV infection in sera samples received throughout the country during 2013 where the IgM anti-HAV test was required by the Clinician. Materials and Methods A specific RT-PCR for open reading frame 2 (ORF2) was used to detect RNA-HEV in 422 sera. The amplified products corresponding to ORF2 were purified, sequenced and phylogenetically analyzed using MEGA6 software program. Results RNA-HEV was detected in 8.53% (36/422) of the samples. The highest rate of positivity was identified in the Western region of the country, specifically in Havana 5.45% (23/422). IgM anti-HAV was detected in 5.21% (22/422) and simultaneous detection of both HAV and HEV was found in 13.88% (5/36) of the samples. The results showed a higher incidence of HEV with respect to HAV (8.53% vs 5.21%) and phylogenetic analysis showed the circulation of genotype 1, subgenotype 1d of the HEV. Conclusions This study corroborated the endemicity of HEV and for the first time the subgenotype 1d, the African variant strain associated to outbreak of hepatitis E, is reported in viral hepatitis cases in Cuba.

Seroprevalence of Hepatitis A and E Virus Infections in Patients with Acute Viral Hepatitis in Hyderabad, India - A One Year Study.

Introduction: Acute viral hepatitis due to faeco- oral group of hepatitis viruses is endemic in India. Outbreaks of hepatitis E virus infection are more common than the hepatitis A virus. Aim: The present study aimed at determining the seroprevalence of IgM antibodies to hepatitis A and E virus in clinically diagnosed acute viral hepatitis cases. And to determine its usefulness against the disease prevention. Study Design: A cross sectional study was carried out on symptomatic patients referred from paediatric and gastro enterology department of Princess Esra hospital. Place and Duration of Study: Around one hundred and eight subjects were analyzed for anti IgM antibodies to hepatitis A and E virus and liver function test in the department of laboratory medicine for Microbiology and biochemistry at Princess Esra Hospital, Deccan College of Medical Sciences between January 2013 and January 2014. Methodology: Blood samples were collected under strict aseptic precautions and tested for anti-HAV and Anti-HEV IgM antibodies using capture elisa from diasorin. Biochemical analysis included estimation of serum aminotransferases, alkaline phosphatase and bilirubin levels. Results: An overall seropositivity of 54% was observed in the present study. More number of the subjects as 46.29% tested positive for anti HAV IgM than for anti HEV IgM as 7.4%. Co-infections were not noticed. Acute viral hepatitis due to hepatitis A virus is more common in children in the age group 6-10 years followed by 11-15 years and lastly 0-5 years indicating the epidemiological shift. Infection with hepatitis E virus was common in adolescent and adults. Males were more susceptible to both the infections than females. Liver function test results correlated well with viral markers indicating damage to the liver parenchyma. The socioeconomic status of the individuals revealed that 95% of the subjects were below poverty line and didn’t have access to proper drinking water and sanitary facilities. None were vaccinated against Hepatitis A virus. Conclusion: The present data suggest that we need to have a dual pronged approach against prevention of acute viral hepatitis caused by A and E virus. Government authorities should prioritize on bringing a uniform improvement in the living standards of the society and make vaccine available to the high risk group at a subsidized rate.

Hepatitis A virus vaccination strategy and pre-immunization screening of Bangladeshi children.

Background: HAV infection is endemic in many developing countries like India, Pakistan, Nepal etc. Several seroprevalence studies show high rates of sero-positivity among children by sub-clinical infection. Therefore mass vaccination against HAV has not been recommended in endemic countries. Objective: To determine whether routine hepatitis A vaccination is indicated for all Bangladeshi children & also to know whether pre-vaccination screening is necessary. Materials & Methods: Serum samples from 254 children aged between 1-15 years were tested for antibody (IgM & IgG) against hepatitis A virus (HAV) to determine the seroprevalence of HAV antibody and do a cost-benefit analysis for decision making about vaccination against HAV among the children of Bangladesh. Results: Hepatitis A virus antibody was positive in 141 (55.5%) of 254 children. Age-specific sero-prevalence was 13 (23.2%) of 56 in 1-3 year,64 (55.2%) of 116 in 3-5 year, 39 (70.9%) of 55 in 5-10 year & 25 (92.6%) of 27 in 10-15 year age group. Cost benefit analysis showed that the total cost of screening followed by vaccination was almost 1.8 times less than the total cost of vaccination of all children without screening. Conclusions: Majority of the children were found sero-positive against HAV around 15 year of age. Therefore mass vaccination against HAV may not be required for Bangladeshi children.

Changes in the seroprevalence of IgG anti-hepatitis A virus between 2001 and 2013: experience at a single center in Korea

BACKGROUND/AIMS: The incidence of symptomatic hepatitis A reportedly increased among 20- to 40-year-old Korean during the late 2000s. Vaccination against hepatitis A was commenced in the late 1990s and was extended to children aged <10 years. In the present study we analyzed the changes in the seroprevalence of IgG anti-hepatitis A virus (HAV) over the past 13 years. METHODS: Overall, 4903 subjects who visited our hospital between January 2001 and December 2013 were studied. The seroprevalence of IgG anti-HAV was analyzed according to age and sex. In addition, the seroprevalence of IgG anti-HAV was compared among 12 age groups and among the following time periods: early 2000s (2001-2003), mid-to-late 2000s (2006-2008), and early 2010s (2011-2013). The chi-square test for trend was used for statistical analysis. RESULTS: The seroprevalence of IgG anti-HAV did not differ significantly between the sexes. Furthermore, compared to the seroprevalence of IgG anti-HAV in the early 2000s and mid-to-late 2000s, that in the early 2010s was markedly increased among individuals aged 1-14 years and decreased among those aged 25-44 years (P<0.01). We also found that the seroprevalence of IgG anti-HAV in individuals aged 25-44 years in the early 2010s was lower than that in the early 2000s and mid-to-late 2000s. CONCLUSIONS: The number of symptomatic HAV infection cases in Korea is decreasing, but the seroprevalence of IgG anti-HAV is low in the active population.

Prevalence of hepatitis A virus in sea food in Iran / Prevalência de vírus da hepatite A em frutos do mar no Irã

The objective of this study was to determine the prevalence of Hepatitis A Virus (HAV) in sea food samples in the Isfahan and Shahrekord townships in Iran. From September 2010 to April 2011, a total of 300 samples of fresh fish, shrimp, crab and lobster were obtained from randomly selected retail stores in the Isfahan and Shahrekord townships in Iran. The samples were tested for the presence of HAV using a reverse transcriptase- polymerase chain reaction method. Out of the total number of samples examined, 8 (2.7%) were found to be positive for HAV. This virus was detected in 5% and 1.7% of fresh fish and shrimp, respectively. This study shows the importance of sea food as potential sources of HAV infection in people in Iran.

O objetivo deste estudo foi determinar a prevalência do vírus Hepatitis A (HAV) em amostras de frutos do mar nas cidades de Isfahan e Shahrekord no Iran. De setembro de 2010 a Abril de 2011 um total de 300 amostras de peixe fresco, camarão, caranguejo e lagosta foram obtidas de lojas de varejo aleatoriamente escolhidas nas cidades de Isfahan e Shahrekord no Iran. As amostras foram testadas para presença de HAV usando o método de reação em cadeia em transcriptase reversa. Do total de amostras examinadas, 8 (2.7%) foram positivas para HAV. Este vírus foi detectado em 5% e 1.7% de peixe fresco e camarão, respectivamente. Este estudo mostrou a importância de frutos do mar como fontes potenciais de infecção HAV em pessoas no Iran.

Seroprevalence of sporadic acute viral hepatitis with their co-infection at tertiary care hospital.

Background: Viral hepatitis is one of the major causes of mortality & morbidity in developing countries. Present study was aimed to know the seroprevalence and co infection of the acute viral hepatitis caused by hepatitis A, hepatitis B, hepatitis C and hepatitis E virus from patient attending one of the largest tertiary care hospitals. Material and Methods: Study was carried out from May 2009 to June 2010 at our hospital. A total of 556 serum samples were tested for HBsAg, antibody for HCV, IgM antibody of HAV and HEV by ELISA method from the patients having clinical signs & symptoms of acute viral hepatitis. Statistical analysis: Chi square test was done and the association was considered to be statistically significant if p < 0.05. Results: Out of 556 suspected cases of acute hepatitis 281 were positive. Among the total positive cases, IgM anti HAV antibodies was 70 (24.91%), IgM anti HEV antibodies was 141 (50.18 %), HBsAg was 65 (23.13%) and HCV was 5 (1.78 %). Dual infection was seen in 14 (4.98 %) cases with 6 children cases. Among the HAV positive patients, 80% were children, In contrast to that HEV infection (63.1%) and HBsAg infection (61.5%) was common in young adult. HCV infection was found common in adults more than 20 years of age. Overall male was affected more than female. Conclusion: Sporadic HEV infection constitutes significant cause of the acute viral hepatitis. In the light of this result a nationwide survey is recommended to confirm this pattern in the other areas. As vaccine for HEV is not available, prevention in form of improvement of socio - economic and hygienic standards of the population is the better option.

Clinical Factors Associated with Hepatitis A Virus Seropositivity in HIV-Infected Adults Living in a Country with an Epidemiologic Shift for Hepatitis A Virus Infection

The number of HIV-infected individuals susceptible to Hepatitis A virus (HAV) infection is increasing in Korea; however, it has proven difficult to devise a vaccination policy therefore because limited seroepidemiologic data exists for them. Accordingly, anti-HAV IgG was measured in 188 HIV-infected adults between July 2008 and July 2010. The nadir CD4+ T lymphocyte counts were not different between the HAV-positive and -negative groups (197 +/- 138 vs 202 +/- 129, P = 0.821). The only factor independently associated with seropositive status was age under 40 yr old (OR 0.017, P < 0.001). Our findings suggest that HAV vaccination in HIV-infected adults should be targeted at persons under the age of 40 yr.

Optimal time for repeating the IgM anti-hepatitis A virus antibody test in acute hepatitis A patients with a negative initial test / 대한간학회지

BACKGROUND/AIMS: The nonspecific clinical presentation of acute hepatitis A (AHA) mandates the detection of anti-hepatitis A virus IgM antibodies (IgM anti-HAV) in the serum for obtaining a definitive diagnosis. However, IgM anti-HAV might not be present during the early phase of the disease. The aim of this study was to determine the optimal time for repeating the IgM anti-HAV test (HAV test) in AHA patients with a negative initial test. METHODS: In total, 261 patients hospitalized with AHA were enrolled for this retrospective study. AHA was diagnosed when the test for IgM anti-HAV was positive and the serum alanine aminotransferase (ALT) level was > or =400 IU/L. Repeat HAV test was conducted after 1-2 weeks if the initial HAV test was negative but AHA was still clinically suspected. RESULTS: The results of the initial HAV test were negative in 28 (10.7%) patients. The intervals from symptom onset to the initial-HAV-test day and from the peak-ALT day to the initial-HAV-test day were significantly shorter in the negative-initial-HAV-test group, but on multivariate analysis only the latter was significantly associated with negative results for the initial HAV test (beta=-0.978; odds ratio [95% confidence interval]=0.376 [0.189-0.747]; P=0.005). The HAV test was positive in all patients when it was performed at least 2 days after the peak-ALT day. CONCLUSIONS: The results of HAV tests were significantly associated with the interval from the peak-ALT day to the HAV-test day. The optimal time for repeating the HAV test in clinically suspicious AHA patients with a negative initial HAV test appears to be at least 2 days after the peak-ALT day.

Enhancement of Protein Productivity of Recombinant Hepatitis A Virus VP1 in Stably Transfected Drosophila melanogaster S2 Cells

The effect of DMSO and sodium butyrate on the production of recombinant hepatitis A virus (HAV) capsid protein VP1 was evaluated and optimized in the culture of stably transfected Drosophila melanogaster S2 cells using culture plates and spinner flasks. The effect of DMSO and sodium butyrate was also evaluated to improve the recombinant VP1 production in stably transfected Drosophila S2 cells. A production level of 0.88 mg of recombinant VP1/liter was obtained in the culture-plate culture of stably transfected S2 cells at 6 days after induction with 0.5 mM CuSO4. The supplements of 2% DMSO and 10 mM sodium butyrate at 4 days post-inoculation increased recombinant VP1 accumulation by 141 and 104%, respectively, resulting in 2.17 and 1.7 mg/liter of recombinant VP1 production. In spinner flasks, recombinant VP1 production reached maximum level at 9 days after induction with 0.5 mM CuSO4, with approximately 4.96 mg/liter of recombinant VP1 production level. When 2% DMSO or 10 mM sodium butyrate was added at 5 days post-inoculation, the recombinant VP1 production was increased to 8.35 and 5.85 mg/liter, respectively. However, the synergistic effects of DMSO and sodium butyrate were not observed. These results indicate that DMSO and/or sodium butyrate can be successfully used to improve the recombinant HAV VP1 production in culture plates and spinner flasks.

Cost-utility analysis on universal childhood hepatitis A vaccination in regions with different anti-HAV prevalence rates of China / 中华流行病学杂志

Objective To explore the inputs and outputs of areas with different anti-HAY prevalence rates on universal childhood vaccination,and to provide a scientific basis for the formulation of the immunization strategy.Methods Since hepatitis A vaccination was scheduled at 12 and 18 months of age for all the healthy children,a single cohort including 1 000 000 individuals was formed in 2009,using the Chinese inactivated vaccine.Decision analysis was used to build Markov-decision tree model.The universal childhood hepatitis A vaccination was compared with nonvaccination group to evaluate the number of symptomatic infection,hospitalization,death,qualityadjusted life years (QALYs) lost,and the incremental cost-utility from the health system and the societal perspectives.Outcomes of the vaccination for the next 70 years were also predicted.The process of analysis was run separately in five regions defined by the anti-HAV prevalence rates (around 50％,50％-69％,70％-79％,80％-89％ and ＞90％ ).Sensitivity analysis was performed to test the stability or reliability of the results,and to identify sensitive variables.Results The study projected that,in the lowest,lower,and intermediate infection regions,the cost and output indicators of universal childhood hepatitis A vaccination were all lower than non-vaccinated group.Universal vaccination could gain QALYs and save both costs from the health systen or the society.In the regions with higher infection rate,the output indicators of universal childhood hepatitis A vaccination were lower than in those non-vaccinated groups,except for the number of death due to hepatitis A,which had a 20 cases of increase.The model also predicted that in the highest infected region,universal vaccination would increase 4 560 814 and 5 840 430 RMB Yuan in the total costs from both the health system and the societies,respectively,when compared to the non-vaccination groups.Universal vaccination would also decrease the numbers of symptomatic infection,hospitalization,and QALYs lost,but would increase 51 deaths due to hepatitis A,and 1507,1929 more RMB Yuan for each QALY gained from the health system and societal respectively,in the regions with highest infection rate.Sensitivity analyses discovered that the infection rate among those susceptible population and the proportion of those who initially under protection but subsequently lost their immunity every year,were the two main sensitive variables in the model.Conclusion Our research discovered that the universal vaccination strategy should be based on the protective period of the vaccine and the anti-HAV prevalence in different endemic areas.