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.

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.

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

HPV B19, HAV and HEV infection among voluntary blood donors in Nantong / 中国输血杂志

【Objective】 To evaluate the viability of nucleic acid test(NAT) of human parvovirus B19 (HPV B19), hepatitis A virus (HAV) and hepatitis E virus (HEV) along with routine NAT for voluntary blood donors, and to analyze HPV B19, HAV and HEV prevalence in Nantong, so as to provide reference for rational blood screening programs. 【Methods】 HPV B19 DNA, HAV RNA and HEV RNA of blood donors in Nantong from November 2021 to May 2022 were detected using NAT, and serological antibody testing was performed on NAT reactive samples. 【Results】 Three HPV B19 DNA was yielded out of 3 440 blood donors, with a positive rate of 0.09%, among which 2 were negative for HPV B19-IgM and 1 was undetermined due to insufficient sample size. HAV RNA and HEV RNA were not detected in 3 440 blood donors. HPV B19, HAV, and HEV NAT were conducted simultaneously with routine HBV, HCV and HIV screening, prolonging the test reports by 20 minutes. 【Conclusion】 Although the HPV B19 DNA, HAV RNA and HEV RNA prevalence among voluntary blood donors in Nantong is low, the risk of transfusion transmitted infection still exsits and can be reduced by NAT.

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.

Economic evaluation of hepatitis A vaccines by income level of the country: A systematic review

Background & objectives: Although several reviews of economic evaluation (EE) studies on hepatitis A virus (HAV) vaccine exist, there remains a need to corroborate such data from time to time. This study aimed to systematically review the literature for reports on EE of HAV vaccination by type of population, characteristics of intervention and income level of the country. Methods: PubMed and Scopus were searched to identify relevant studies from inception up to May 2021 using topic-specific key words in various combinaiton. Full EE studies comparing HAV vaccination to no vaccine or immunoglobulin were included. The risk of bias was assessed by using the ECOBIAS checklist. Results: Among the 1984 identified studies, 43 were found eligible. Of these, 27 were from high-income countries (HICs), 15 from middle-income countries (MICs), and one from low income country. Majority of the studies used Markov model and/or decision tree (n=26). Eight studies used a dynamic model. The discount rate, perspective and time horizon varied across the studies. Universal HAV vaccination without screening was cost-effective among children (14/16, 87.5%) and adolescents (1/5, 20%) but not in adults (0/4, 0%). Analysis by the level of income found that universal HAV vaccination among children without screening was cost-effective in 81.8 per cent of the studies conducted in MICs (9/11) as compared to 66.7 per cent in HICs (4/6). About one-third of the studies conducted among children found that screening and HAV vaccination were cost-effective compared to no vaccination. Interpretation & conclusions: The finding of this review suggest that universal vaccination of children without screening was likely to be cost-effective, especially in MICs. Nevertheless, it should be noted that the methodology varied across studies. Several aspects should also be considered in transferring the EE results across jurisdictions.

Actualización sobre hepatitis A

Introducción: La hepatitis es una enfermedad inflamatoria que afecta al hígado. El virus de la hepatitis A produce un cuadro infeccioso agudo por lo general autolimitado en el ser humano, para el cual no existe tratamiento específico. Objetivo: Realizar una actualización acerca del comportamiento y manejo actual de la infección por el virus de la hepatitis A. Métodos: Se realizó una revisión bibliográfica durante septiembre y octubre de 2021, se consultaron 156 artículos en idioma inglés y español de bases de datos como SciELO, PubMed, LILACS y Medigraphic, de ellos se utilizaron 30 citas. Se empleó la combinación de términos y operadores booleanos y métodos teóricos como: análisis-síntesis, inducción-deducción e histórico-lógico. Resultados: Se abordó el comportamiento de la infección causada por el virus de la hepatitis A, se hizo hincapié en su epidemiología, características morfológicas y funcionales que permiten su infectividad, evolución y manifestaciones clínicas, complicaciones, tratamiento y perspectivas actuales. Conclusiones: El virus de la hepatitis A produce un cuadro infeccioso agudo. Se presenta tanto de forma esporádica como epidémica. Es la forma más común de hepatitis viral aguda. La transmisión es de persona a persona por vía fecal oral. Los síntomas relacionados con las hepatitis virales agudas son variables e inespecíficos. El diagnóstico de certeza se realiza mediante las pruebas serológicas y técnicas de biología molecular para la identificación del virus. No existe tratamiento específico para esta infección.

Introduction: Hepatitis is an inflammatory disease that affects the liver. The hepatitis A virus produces a generally self-limited acute infectious picture in humans, for which there is no specific treatment. Objective: To update on current behavior and management of hepatitis A virus infection. Methods: A bibliographic review was carried out during September and October 2021, consulting 156 articles in English and Spanish from databases such as SciELO, PubMed, Lilacs and Medigraphic, of which 30 citations were used. The combination of Boolean terms and operators and theoretical methods such as: analysis-synthesis, induction-deduction and historical-logical was used. Results: The behavior of the infection caused by the hepatitis A virus was addressed, emphasizing its epidemiology, morphological and functional characteristics that allow its infectivity, evolution and clinical manifestations, complications, treatment and current perspectives. Conclusions: The hepatitis A virus produces an acute infectious picture. It occurs both sporadically and epidemically. It is the most common form of acute viral hepatitis. Transmission is person-to-person by the fecal oral route. Symptoms related to acute viral hepatitis are variable and nonspecific. The diagnosis of certainty is carried out by means of serological tests and molecular biology techniques for the identification of the virus. There is no specific treatment for this infection.

Incidence of hepatitis A and hepatitis E viruses and norovirus and rotavirus in fish and shrimp samples caught from the Persian Gulf / [Incidência de vírus da hepatite A e hepatite E e norovírus e rotavírus em amostras de peixes e camarões capturados no Golfo Pérsico]

Foodborne viruses including hepatitis A virus (HAV), norovirus (NoV), rotavirus (RoV) and hepatitis E virus (HEV) are easily transmitted through contaminated seafoods. The current research was done to assess the incidence of RoV, NoV GI and GII,hAV and hEV in fish and shrimp samples caught from the Persian Gulf, Iran. Three-hundred and twenty fish and shrimp samples were collected. The presence of foodborne viruses were assessed by the real-time PCR. Forty-nine out of 320 (15.31%) fish and shrimp samples were positive for foodborne viruses. Distribution of hAV, NoV GI and NoV GII amongst all studied samples were 0.93%, 5.93% and 8.43%, respectively. hEV and RoV viruses were not found in studied samples. Parastromateus niger and Scomberomorus commerson fish and Penaeus monodon shrimp were the most frequently contaminated samples. Simultaneous incidence of hAV and NoV GI and hAV and NoV GII were 0.31% and 0.93%, respectively. Distribution of foodborne viruses in samples collected through spring, summer, autumn and winter seasons were 14.28%, 9.33%, 11.76% and 24.44%, respectively. Findings revealed that the incidence of foodborne viruses was significantly associated with seafood species and also season of sampling.(AU)

Vírus transmitidos por alimentos, incluindo hepatite A (HAV), norovírus (NoV), rotavírus (RoV) e hepatite E (HEV) são facilmente transmitidos através de frutos do mar contaminados. Esta pesquisa foi realizada para avaliar a incidência de RoV, NoV GI e GII, hAV e hEV em amostras de peixes e camarões capturadas no Golfo Pérsico, Irã. Foram coletadas 300 amostras de peixes e camarões. A presença de vírus transmitidos por alimentos foi avaliada por PCR em tempo real. Quarenta e nove das 320 amostras de peixes e camarões (15,31%) foram positivas para vírus transmitidos por alimentos. A distribuição de hAV, NoV GI e NoV GII entre as amostras estudadas foi 0,93%, 5,93% e 8,43%, respectivamente. Os vírus hEV e RoV não foram encontrados nas amostras estudadas. Os peixes Parastromateus niger e Scomberomorus commerson e o camarão Penaeus monodon foram as amostras mais frequentemente contaminadas. A incidência simultânea de hAV e NoV GI, e hAV e NoV GII foi de 0,31% e 0,93%, respectivamente. A distribuição dos vírus transmitidos por alimentos nas amostras coletadas na primavera, verão, outono e inverno foi de 14,28%, 9,33%, 11,76% e 24,44%, respectivamente. Os resultados demonstram que a incidência de vírus transmitidos por alimentos foi significativamente associada às espécies de frutos do mar e também à época da amostragem.(AU)

Old dietary cultural practices leading to suboptimal nutritional intake of calories in children with acute viral hepatitis

Background: Acute viral hepatitis (AVH) is one of the common infections in developing nation. Hepatotrophic virus causes AVH and most of the time it has self-limiting course. HAV is most common cause of AVH. Suboptimal dietary cultural practices are common in our nation. This study was done to know the impact of suboptimal dietary practices done in AVH.Methods: Observational study conducted with 50 children with AVH. Dietary assessment was done at three point of time.  First During AVH, second, pre illness and then two weeks after nutritional counselling. Data of dietary modification done by parents/caregiver during AVH recorded. History of herbal medicine taken for treatment of AVH enquired.Results: HAV positive was (72%), HEV positivity was in 8 (16%) cases, hepatitis B was positive in 4 (8%) children and dual infection HAV+HEV was detected in 2 (4%) children. Many parents did dietary modification due to AVH and also by some local treating physicians, 18 (36%) patients were also on herbal medication for treatment of AVH. During AVH the mean calorie and protein requirement decreased from 96% to 54% and 92% to 43% respectively. Two weeks after nutritional counseling the mean calorie and protein intake increased from 54% to 88% and 43% to 84% respectively. This change in calorie and protein was statistically significant (p<0.01) Non-vegetarian diet preference was restarted in many of them after nutritional counseling.Conclusions: This observational study revealed that suboptimal dietary cultural practices are prevalent in India. Majority of children were on bland diet and receiving inadequate proteins and calories.

Status of adult immunity to hepatitis A virus in healthcare workers from a tertiary care hospital in north India

Background & objectives: Humans are considered to be the principal host for hepatitis A virus (HAV) infection. In India, heterogeneous groups of susceptible individuals coexist in different regions. There has been a decline in antibody titres to HAV among young adults which may pose a major public health problem. The objective of this study was to assess the IgG anti-HAV level among healthcare workers (HCWs) in the age group of 20-60 yr and its association with the socio-demographic variables. Methods: Blood sample (2 ml) was collected under aseptic conditions from each participant followed by the preparation of serum and storing at ?20癈. ELISA-based kits were used for the determination of IgG antibodies to HAV in the human serum samples. Results: Two hundred and fifty four HCWs were enrolled. IgG anti-HAV antibodies were detected in 97.2 per cent of the samples analyzed. No differences were observed in the levels of IgG anti-HAV antibody and education, income, occupation and socio-economic classes of the HCWs. A seropositivity rate of over 90 per cent was seen amongst all the socio-economic classes. Interpretation & conclusions: High levels of IgG protective antibodies were seen among the studied HCWs, hence HAV vaccination may not be required. It will be advisable to do a cost-benefit analysis of vaccination for HAV.

Prevalence of hepatitis A virus in patients attending a referral hospital in Bubanza Province, Northwest Burundi

Background: Viral hepatitis is a public health problem wide world. Hepatitis A, transmitted by fecal-oral route, is an infectious viral disease caused by hepatitis A virus and mainly due to poor sanitation. This study aimed to determine the prevalence of hepatitis A virus and associated factors in patients attending Mpanda referral hospital in Northwest Burundi.Methods: A cross-sectional study was done from November 2017 to January 2018 on 385 participants aged 2 years and above. Participants were recruited using a systematic random sampling technique. Data were collected using questionnaire from consented/assented participants. Five millilitres of venous blood was collected and analyzed. Anti-hepatitis A virus antibodies were screened using Enzyme Immuno Assay. Data were analyzed using Statistical Package for the Social Sciences version 16.0 software. A descriptive analysis was followed by bivariate analysis using a Chi-square test for comparison of various sub-groups with 5% statistical significance level. Odds ratio and 95% Confidence Intervals were calculated and presented.Results: The median age of the participants was 23 years and the range 72 years. The overall prevalence of Hepatitis A virus was 60.3%. There was a significant association between age [OR=7.22 (4.04-12.93), P <0.001], lack of clean water [OR=10.07 (5.63-18.01), P <0.001], traditional latrines [OR=1.86 (1.02-3.40), P=0.04] and Hepatitis A Virus seroprevalence.Conclusions: Present study shows high prevalence of HAV infection in patients attending Mpanda Referral Hospital. Younger age, lack of clean water and traditional latrines play roles in increasing prevalence of HAV infection in both rural and urban areas.

Concentration of hepatitis A virus from mimicked water samples by membrane filtration method / 中华实验和临床病毒学杂志

Objective@#To optimize the membrane filtration method for hepatitis A virus concentration from mimicked water samples.@*Methods@#Mimicked water samples containing HAV particles were prepared and concentrated by positively charged membrane and negatively charged membrane respectively. Then different method including direct lysis, shaker, vortex and ultrasonication were used to elute HAV followed by the quantification of HAV by Taqman Real-time RT-PCR. The data were analyzed by professional statistical software.@*Results@#In the present study, when mimicked water samples contained 300 TCID50 of HAV, there was no significant difference between the concentration effects by negatively charged membrane and positively charged membrane (P=0.825>0.05). However, when HAV in mimic water samples was up to 1500 TCID50 and 7500 TCID50, the recycle efficiency by negatively charged membrane was higher than that by positively charged membrane (P<0.01). Additionally, this study found that HAV recycle ratio could be up to (68.17±16.79)% when direct lysis was used for viral elution, therefore direct lysis was proved much better than shaker, vortex and ultrasonication. They were significantly different (P<0.0001).@*Conclusions@#Elution process played the key role in HAV concentration when membrane filtration method was used. Direct lysis was proved much better than other method and it was the most efficient way in HAV recycle from mimicked water samples.

Causes and countermeasures for repeated outbreaks of hepatitis A among adults in Korea / 한국역학회지

The 2019 hepatitis A outbreak has become increasingly prevalent among adults in Korea and is the largest outbreak since that in 2009–2010. The incidence in the current outbreak is highest among adults aged 35–44 years, corresponding to the peak incidence among those aged 25–34 years 10 years ago. This may indicate a cohort effect in the corresponding age group. Causes of these repeated outbreaks of hepatitis A in Korea are low level of immunity among adults, Korean food culture that consumes raw seafood such as salted clam and inadequate public health system. Among countermeasures, along with general infectious disease control measures including control of the infectious agent, infection spread, and host, urgent actions are needed to review the vaccination policy and establish an adequate public health system.

Metagenomic analysis reveals Hepatitis A virus in suspected yellow fever cases in Brazil

Using a metagenomic approach, we identified hepatitis A virus among cases of acute febrile illnesses that occurred in 2008-2012 in Brazil suspected as yellow fever. These findings reinforce the challenge facing routine clinical diagnosis in complex epidemiological scenarios.

A Case of Graves' Disease Accompanied with Acute Hepatitis A Virus Infection / 대한소화기학회지

Concurrent presentation of acute hepatitis A virus (HAV) infection and Graves' disease has not been reported in literature worldwide. Although there is no well-established mechanism that explains the induction of Graves' disease by HAV to date, our case suggests that HAV infection may be responsible for inducing Graves' disease. A healthy 27-year-old female presented fever, palpitation, and diarrhea, and she was subsequently diagnosed as acute HAV infection. Concurrently, she showed hyperthyroidism, and the diagnosis was made as Graves' disease. She had never had symptoms that suggested hyperthyroidism, and previous thyroid function test was normal. Acute HAV infection was recovered by conservative management, however, thyroid dysfunction was maintained even after normalization of liver enzymes. Methimazole was used to treat Graves' disease. We report a case of concurrent acute HAV infection and Graves' disease in a patient without preexisting thyroid disease. This suggests that HAV infection may be a trigger for an autoimmune thyroid disease in susceptible individuals.

Whole-genome and quasispecies sequences analysis of three hepatitis A virus isolates in China / 中华实验和临床病毒学杂志

Objective@#To analyze the genetic characteristics of whole-genome and quasispecies sequences from three hepatitis A virus (HAV) strains in China.@*Methods@#Serum samples from acute hepatitis A patients were collected and viral RNA extraction, transcription, nested PCR, sequencing and assembling were performed to gain near full-length sequences; cloning-based sequencing of the full-length VP1-2 A region was also performed.@*Results@#Genotyping showed that the nucleotide and amino acid identities among three strains on VP1-2 A junction region were both 100% and all belonged to subgenotype IA; the nucleotide and amino acid identities on whole-genome region were 99.9%-100% and 100% respectively, and shared the highest identities with AH2 strain from GenBank of 98.5% in nucleotide and 99.7% in amino acid level; no amino acid variation was found among published neutralizing antigenic sites. Within cloning sequences from each strain, the nucleotide and amino acid identities were 99.0%-100% and 98.1%-100%, while among all cloning sequences were 99.0%-100% and 97.2%-100%. The variation rate of nucleotide and amino acid in VP1-2 A junction region were both higher than that of partial VP1 region.@*Conclusions@#Sequences among three strains in VP1-2 A region were identical, the nucleotide and amino acid identities in both whole-genome region and among quasispecies sequences were relatively high to deduce that they were from the same outbreak. This study provides new insight for identification of HAV transmissions and tracing investigations.

Successive subculture and genetic stability of hepatitis A virus vaccine strain H2 / 中华实验和临床病毒学杂志

Objective@#To investigate the genetic stability of virus seed H2M20K7 (K7) of live attenuated Hepatitis A virus H2 strain (HAV, H2 strain) for production of hepatitis A (Live) vaccine, lyophilized after continuous passages.@*Methods@#The virus seed K7 of H2 strain was proliferated and passaged in KMB17 cells in cell factories. Viruses of different passages were harvested after continuous passages. Virus RNA was extracted and the complete genomes of different virus passages (K7, K10, K11, K13, K15, K18) were sequenced by using next-generation deep sequencing. The mutation rates of different passages were compared. The infectivity titers of different virus passages of H2 strain were tested by ELISA.@*Results@#The mutation rates of complete genomes of different passages were low after continuous passages of master virus seed. The structure of gene was stable and non-synonymous mutation rate was lower than 0.57%. The mutation rate of 5 ’non-coding regions was lower than 0.1%. There was no significant mutation in VP1/2 A and 2C virulence site. The infectious titers of H2 strains of different passages were within 7.76-8.50 lgCCID50/ml. No statistically significant difference was found in this study.@*Conclusions@#The gene structure of the master virus seed, working seed and different passages of H2M20K7 after subculture was stable and the mutation rate was low. No significant mutation was found in 5’non-coding regions, and the critical virulence sites such as VP1/2 A, 2B and 2C showed attenuated characteristics with low mutation rate. Virulence of the virus did not changed. The H2 strain maintained stable viral infectivity and genetic stability and comply with the requirements as virus seed for vaccine manufacturing.

Virus hepatitis E: presentación hepática y encefálica / Hepatitis E virus: hepatic and encephalic presentation / Vírus da hepatite E: apresentação hepática e encefálica

Resumen: La infección por el virus de la hepatitis E se presenta en la mayoría de los casos como cuadros autolimitados indistinguibles de una hepatitis a virus A. En determinados escenarios clínicos(embarazadas y pacientes con enfermedad hepática crónica) pueden determinar cuadros graves. Las manifestaciones extra hepáticas de la enfermedad son excepcionales, y en particular las neurológicas. Presentamos un caso de encefalitis aguda como manifestación de una hepatitis a virus E.

Abstract: The Hepatitis E virus infection is usually an acute self-limited disease, but in some clinical situations (pregnancy and chronic liver desease) may presented as failure. The extra hepatic manifestation are not common in particular de neurological ones. We describe a case with acute encephalitis as a presentation of a hepatitis E virus infection.

Resumo: Na maioria dos casos, a infecção do vírus da hepatite E é uma condição auto-limitante indistinguível do vírus da hepatite A. Em certas situações clínicas (mulheres grávidas e pacientes com doença hepática crônica), condições severas podem ser determinadas. As manifestações extra-hepáticas da doença são excepcionais e, em particular, neurológicas. Apresentamos um caso de encefalite aguda como manifestação do vírus da hepatite E.

Prevalence of hepatitis A antibody among population covered by different hepatitis A immunization strategies in Shandong Province, 2015, China / 中华预防医学杂志

Objective@#To evaluate prevalence of hepatitis A antibody (anti-HAV IgG) among population covered by different hepatitis A vaccine immunization strategies in Shandong Province in 2015.@*Methods@#In October 2015, according to the geographical location of Shandong Province, the stratified random sampling method was used to stratify the 17 municipal distrcts, and the random number table method was used for sampling, First, two eastern cites (Qingdao, Rizhao), two western cities (Liaocheng, Zaozhuang) and three central cities (Jinan, Zibo and Laiwu) were selected; secondly, one county was drawn from each city. Finally, the participants were divided into five age groups including ≤7 years (age group covered by free hepatitis A vaccination strategy), 8-11 years (age group who receive hepatitis A vaccination at their own charge), 12-24 years (age group covered by catch-up vaccination of hepatitis A), 25-34 years (age group born before hepatitis A vaccine was used) and ≥35 years (age group born before hepatitis A vaccine was used). After all the paticipants or their guardians asked and registered basic information such as age, gender, home address, blood samples were collected from them and anti-HAV IgG was detected by ELISA method. The positive rate of anti-HAV IgG and 95%CI were calculated.@*Results@#A total of 1 654 participants were involved in the final analysis, including 856 males (51.75%) and 798 females (48.25%) whose mean age was (13.44±13.06) years. The crude positive rate of anti-HAV IgG was 91.41% (1 512/1 654, 95%CI: 89.96%-92.72%) and the age-adjusted rate was 90.93% (95%CI: 90.92%-90.94%). The positive rates of anti-HAV IgG was at the highest level in the age group of ≤7 years (95.90%, 95%CI: 95.88%-95.91%) and was at the lowest level in the age group of 25-34 years (83.23%, 95%CI: 83.21%-83.25%). The age-specific positive rates of anti-HAV IgG in eastern areas (96.79%, 95%CI: 96.78%-96.80%) were higher than those in both middle areas (86.66%, 95%CI: 86.65%-86.67%) and western areas (91.96%, 95%CI: 91.95%-91.97%).@*Conclusion@#The positive rate of anti-HAV IgG was high among the general population in Shandong Province, but relatively low among young and middle-aged adults. Besides the routine immunization of hepatitis A among the children, more efforts should be taken for the prevention and control of hepatitis A among young and middle-aged adults in Shandong Province, especially in central and western areas.

Murine enteroids culture system and Marmota Himalayana Hepatovirus replication by the enteroids / 中华实验和临床病毒学杂志

Objective@#To establish mouse intestinal tissue and its application in the proliferation of Marmota Himalayana hepatovirus (MHHAV).@*Methods@#The intestinal crypts of Balb/C mice were digested with EDTA at a high concentration, cultured in Matrigel and passaged into frozen intestinal enteroids. The monolayers were inoculated on 96-well plates and infected with MHHAV. Real-time qPCR virus concentrations at different time points were compared with those of a number of cell lines routinely used to develop hepatitis A virus.@*Results@#The mouse intestinal tissue culture system was successfully established. MHHAV did not proliferate in the cell lines commonly used for the culture of hepatitis A virus, but proliferated well in the intestinal tissues of mice. The virus multiplied about 3 700 fold in 4 days.@*Conclusions@#Mice intestinal enteroids can be used for the isolation and culture of MHHAV.