Dual infection with hepatitis A and E viruses in outbreaks and in sporadic clinical cases: Cuba 1998-2003

Viral hepatitis ranks as the fifth cause of morbidity for infectious diseases in Cuba. Epidemics are observed frequently in the population, the hepatitis A virus being the main agent responsible for such epidemics. Previous reports also confirmed the circulation of the hepatitis E virus. From 1998 to 2003, 258 serum samples were collected by the Reference Laboratory on Viral Hepatitis during 33 outbreaks of acute viral hepatitis as well as from 39 sporadic clinical cases. Sera were tested for anti-HAV and anti-HEV IgM by EIA. Overall of the 33 outbreaks studied sera from 12 (36.4%) were positive for anti-HAV IgM only, from 7 (21.2%) were positive for anti-HEV IgM only, and from 14 (42.4%) were positive for antibodies to both viruses. Individually of the 258 sera collected, 137 (53.1%) were positives for anti-HAV IgM, 20 (7.8%) were positives for anti-HEV IgM, 33 (12.8%) were positives for both markers and 68 (26.4%) were negative to both. Of the clinical cases, 4 (10.3%) were positives for anti-HAV IgM, 13 (33.3%) were positives for anti-HEV IgM and 5 (12.8%) were positives for both markers. Seventeen (43.6%) sera were negatives for all viral hepatitis markers available (A-E). A high positivity for HEV was found in outbreaks tested with the kit produced by CIGB. In particular HEV seems to infect individuals of all ages. The results demonstrate the co-circulation of and co-infection with two enterically transmitted viruses; however a higher positivity was observed for anti-HAV than to anti-HEV (53.1% vs. 7.8%) in outbreaks.     

Coronavirus infection of rat dorsal root ganglia: ultrastructural characterization of viral replication, transfer, and the early response of satellite cells

Swine hemagglutinating encephalomyelitis virus (HEV) has been shown to have a capability to gain access to the cell bodies of sensory neurons after peripheral inoculation, resulting in ganglionic infection. It is not clearly understood how this virus is replicated within and released from the sensory neurons, and it remains to know how satellite cells response to the HEV invasion. By ultrastructurally examining HEV-infected rat dorsal root ganglia, we found that HEV in the cell bodies of infected neurons budded from endoplasmic reticulum-Golgi intermediate compartments, and were assembled either individually within small vesicles or in groups within large vesicles. The progeny virions were released from the sensory neurons mainly by smooth-surfaced vesicle-mediated secretory pathway, which occurred predominantly at the perikaryal projections and infoldings of sensory neurons. Released HEV particles were subsequently taken up by the adjacent satellite cells. Almost all virus particles in the cytoplasm of satellite cells were contained in groups within vesicles and lysosome-like structures, suggesting that these glial cells may restrict the local diffusion of HEV. These observations give some insights into the pathogenesis of coronavirus infection and are thought to help understand the interactions between sensory neurons and their satellite cells.     

戊型病毒性肝炎实验室诊断研究进展

戊型病毒性肝炎（简称戊型肝炎）在临床表现和实验室生化检查方面与其他病毒性肝炎差别不大,诊断主要依靠实验室病原学检查。传统观念认为,戊型肝炎为自限性疾病,如果患者为慢性肝炎可不考虑是戊型肝炎病毒（HEV）感染。但已有研究发现,     

Hepatitis E virus infection after haploidentical haematopoietic stem cell transplantation: incidence and clinical course

Hepatitis E virus (HEV) is increasingly found to cause hepatitis in allogeneic haematopoietic stem cell transplantation (HSCT) patients. However, little is known about HEV infection in patients receiving haploidentical HSCT (haplo-HSCT). Here, we retrospectively evaluate the incidence and clinical course of HEV infection in haplo-HSCT patients. From January 2014 to July 2017, 177 patients with unexplained elevated transaminases after receiving haplo-HSCT at Peking University Institute of Haematology were screened for HEV using HEV serology. HEV RNA was assessed in blood samples when HEV-IgG and/or IgM antibodies were positive. Acute HEV infection was identified in 7 patients (3·9%), 1 of whom had developed a chronic HEV infection. The median time from haplo-HSCT to HEV infection was 17·5 (range, 6–55) months. HEV infection was confirmed by the presentation of anti-HEV IgM + anti-HEV IgG (rising) (n = 5) or HEV-RNA + anti-HEV IgM + anti-HEV IgG (n = 2). None of the patients died of HEV infection directly: 2 patients with HEV infection died showing signs of ongoing hepatitis, and 5 patients cleared HEV with a median duration of HEV infection of 1·5 (range, 1·0–5·7) months. In conclusion, HEV infection is a rare but serious complication after haplo-HSCT. We recommend screening of HEV in haplo-HSCT.     

Occurrence of water-borne enteric viruses in two settlements based in Eastern Chad: analysis of hepatitis E virus, hepatitis A virus and human adenovirus in water sources

Hepatitis E virus (HEV) is a common cause of water-borne acute hepatitis in areas with poor sanitation. In 2004 an outbreak of HEV infection affected around 2,000 people in Eastern Chad (Dar Sila). This paper describes the decrease in the incidence of acute jaundice syndrome (AJS) from 2004 until 2009 when a mean incidence of 0.48 cases/1,000 people/year was recorded in the region. Outbreaks of AJS were identified in some of the camps in 2007 and 2008. Moreover, water samples from drinking water sources were screened for human adenoviruses considered as viral indicators and for hepatitis A virus and HEV. Screening of faecal samples from donkeys for HEV gave negative results. Some of the samples were also analysed for faecal coliforms showing values before disinfection treatment between 3 and >50 colony forming units per 100 mL. All water samples tested were negative for HEV and HAV; however, the presence of low levels of human adenoviruses in 4 out of 16 samples analysed indicates possible human faecal contamination of groundwater. Consequently, breakdowns in the treatment of drinking water and/or increased excretion of hepatitis viruses, which could be related to the arrival of a new population, could spread future outbreaks through drinking water.     

戊型肝炎病毒感染ZCLA长爪沙鼠的实验研究

目的:确定ZCLA长爪沙鼠作为戊型肝炎病毒感染动物的可能性。方法:选取鉴定含有戊型肝炎病毒的患者粪便,PBS混匀制成10%的粪悬液后10-1原液浓度腹腔接种ZCLA长爪沙鼠100μl,分别收集实验第0,7,14,21,28,35,42,49天的粪便,荧光定量PCR测定接种粪悬液以及收集的粪便所含戊型肝炎病毒滴度。结果:ZCLA长爪沙鼠腹腔接种的粪悬液所含戊型肝炎病毒滴度为4.1×103 copies/g,实验第7天ZCLA长爪沙鼠出现粪便排毒现象并持续至第35天,第7天粪便中戊型肝炎病毒滴度最高,为1.9×103 copies/g,随着实验进程时间推移HEV滴度逐渐降低。呈现下降趋势,第42天和第49天粪便HEV检测均为阴性。对照组粪便HEV检测均为阴性。结论:ZCLA长爪沙鼠能够感染戊型肝炎病毒,可以考虑进一步应用于戊型肝炎病毒动物感染研究。     

抗HEV中和抗体13D8的单链抗体的构建及表达

目的 :降低HEV中和性单抗 (mAb) 13D8的鼠源性 ,表达其单链抗体 (scFv)。方法 :从分泌 13D8鼠mAb的杂交瘤细胞中 ,通过RT PCR克隆mAb的VL、VH 基因 ,并进一步组装成VH linker VL 型的scFv片段。将scFv片段克隆到pTO T7载体中 ,在大肠杆菌中进行表达。用ELISA、Westernblot检测scFv的活性。结果 :SDS PAGE表明 ,13D8的scFv在E .coli中得到高效表达 ,表达量达菌体总蛋白的 2 6 .8%左右 ,表达产物主要以包涵体的形式存在。间接ELISA和Westernblot检测表明 ,表达的 13D8的scFv能与HEVOFR2区中一段重组蛋白(NE2 )特异结合。竞争ELISA表明 ,scFv与原鼠mAb识别的为同一表位。结论 :成功地表达出具有免疫学活性的 13D8的scFv。     

Complete sequence of a Kyrgyzstan swine hepatitis E virus (HEV) isolated from a piglet thought to be experimentally infected with human HEV

Hepatitis E virus (HEV) was identified by RT-PCR amplification with degenerate ORF2 primers in the stool of a piglet experimentally inoculated with a stool suspension from a patient with acute hepatitis during an outbreak of non-A, non-B hepatitis in Kyrgyzstan. Further characterization by sequencing of the complete genome and phylogenetic analysis showed that the piglet isolate was most closely related to HEV genotype 3. Because the original human stool specimen used to inoculate the piglet was no longer available, stool samples from three patients obtained during the same outbreak were sequenced and found to be HEV genotype 1. These findings suggest that the HEV isolated from the swine stool was probably an HEV enzootic in Kyrgyzstan and not the virus inoculated from the human stool.     

A new Chinese isolate of Hepatitis E virus: Comparison with strains recovered from different geographical regions

The full-length cDNA of a new Chinese strain (KS2-87) of Hepatitis E virus (HEV) has been constructed and sequenced. The 5 noncoding region of KS2-87 is 26 nucleotides in length, which is one nucleotide shorter than that of HEV (B1) (Burma) and 23 nucleotides longer than that of HEV (Mexico). Comparison of the nucleotide and amino acid sequences of KS2-87 with all other published HEV sequences showed that KS2-87 was closer to two other Chinese strains (CHT-88, CHT-87) and SAR-55 (Pakistan) than to HEV (B1) and HEV (B2) (Burma) or HEV (Mexico). Comparisons of partial sequences of genes encoding a nonstructural and a structural protein revealed the existence of genetically related groups of HEV within geographical regions, whereas larger nucleotide differences were seen among isolates that were more geographically and epidemiologically distant.Sequence accession No. L25595.     

Prevalence of hepatitis E virus infection in multiple transfused Brazilian patients with thalassemia and sickle cell disease

Hepatitis E virus (HEV) is a leading cause of acute hepatitis worldwide. The virus is acquired by fecal-oral route; however, it can also be transmitted by blood transfusion. The objective of the study was to examine anti-HEV immunoglobulin G and HEV RNA prevalence in multiple transfused patients with thalassemia and sickle cell disease (SCD), and in blood donors. The HEV seroprevalence in the patients was 13% (20% in thalassemics; 7.7% in SCD), and 11% in blood donors. No positive result for HEV RNA was obtained. This is a pioneer study examining HEV circulation in Brazilian patients with hemoglobinopathies.