OBJECTIVETo determine the seasonal prevalence of genotype-IV hepatitis E virus (HEV) in swine herds in Eastern China and explore the phylogenetic relationship between swine HEV and human HEV in the situation that zoonotic features of HEV had been confirmed.

METHODSFrom September 2007 to June 2008, a total of 1200 swine bile specimens were collected from three slaughter houses located in Zhejiang, Anhui and Jiangsu, the Eastern China, and detected for HEV RNA by using nested RT-PCR. The positive PCR products were sequenced. Then the swine HEV were phylogenetically determined with human HEV isolated in Eastern China.

RESULTSThe positive rate for HEV RNA in swine herds was 4.5% totally. Significant differences of HEV detection were not observed among seasonal pattern (Sep - Oct: 6%, Dec - Jan: 4.33%, Mar - Apr: 4.33%, May - Jun: 3.33%) but in geographic distribution (Jiangsu: 6%, Anhui: 5%, Zhejiang: 2.5%). Regardless of isolation from different areas,swine and human genotype-IV HEV shared a high similarity. Phylogenetically, there were 80% - 100% and 96% - 100% identities within swine genotype-lV HEV at the nucleotide and amino acid levels respectively. Between swine HEV and human HEV, there were also similarities of 76% -99% and 95% - 100%. It was noted that some human and swine isolates were clustered with bootstrap values of > 90%.

CONCLUSIONGenotype-IV HEV is widely prevalent in swine herds in Eastern China and original common ancestor of evolution and transmission was implied. The sustaining prevalence within swine herds should have a probable influence on the epidemic situation of hepatitis E in human beings.

Animals ; China ; epidemiology ; Genotype ; Geography ; Hepatitis E ; epidemiology ; Hepatitis E virus ; classification ; genetics ; isolation & purification ; Humans ; Phylogeny ; Prevalence ; Seasons ; Sequence Homology, Nucleic Acid ; Swine ; Swine Diseases ; epidemiology ; genetics ; virology

The primers and probes for the Real-time RT-PCR were designed based on the multiple sequence (swine and humans HEV strains) alignments of the ORF3 region of genotype 4 HEV. A rapid, sensitive and stable TaqMan Real-time RT-PCR assay was established, and its specificity and sensitivity were assessed, and comparison of the Real-time RT-PCR with conventional and nested RT-PCR was performed. The results found that the crossing points showed linearly proportional to the logarithm of the input copy number. The correlation coefficient (R2) and the slope value of the standard curves with plasmid DNA were 0.994 and -3.312, respectively. The efficiency (E) of the PCR was 100%. Coefficients of variation values of the different diluted plasmid DNA were low in the same or different repeated experimental group. In addition, the assay was able to correctly detect genotype 4 HEV RNA from swine fecal samples. The sensitivity of established assay was 100-fold higher than that of conventional RT-PCR and 10-fold higher than nested RT-PCR.
Animals ; DNA Primers ; genetics ; Disease Reservoirs ; virology ; Feces ; virology ; Fluorescence ; Genotype ; Hepatitis E ; virology ; Hepatitis E virus ; classification ; genetics ; isolation & purification ; Humans ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Swine ; virology

OBJECTIVETo determine the genotype and phylogenetic characteristics of hepatitis E virus (HEV) strains isolated from the human and swine in Anqing City.

METHODSTwenty seven sera from sporadic hepatitis E patients and 400 commercial swine bile samples were collected in Anqing City. According to the collection time, the bile samples were equally divided into 4 groups which were named group A, B, C and D respectively. Nested reverse transcription polymerase chain reaction (RT-PCR) and DNA sequencing technology were performed to obtain the DNA sequences of HEV RNA Open Reading Frame 2 (ORF2) (150 nt) for all the serum and bile samples. The sample sequences and prototype sequences from the GenBank were aligned and their nucleotide sequence identities were calculated. A phylogenetic tree constructed according to the Bayesian inference method was used to analyze the genotype and phylogenetic relationship between the human and swine HEV strains isolated in Anqing City.

RESULTSThe male-to-female sex ratio of the patients was 2.86:1 and the average age was 56.78 years old. Sixteen out of 27 serum (59.26%) samples were HEV RNA positive. Human HEV strains isolated in Anqing City shared 74.75% - 82.99%, 75.26% - 83.64%, 72.77% - 80.57% and 88.03%-91.63% nucleotide sequence identities with prototype I, II, III and IV HEV strains respectively. HEV RNA was detected in 22 out of 400 bile samples (5.5%). The swine HEV detection rates for group A, B, C and D were 7.00%, 3.00%, 9.00% and 3.00% respectively, showing no significant difference among these groups (χ(2) = 5.20, P = 0.16). Swine HEV strains isolated in Anqing City shared 75.24% - 83.42%, 75.93% - 84.19%, 72.86% - 80.64% and 88.15% - 91.79% nucleotide sequence identities with prototype I, II, III and IV HEV strains respectively. Phylogenetic analysis revealed that all the HEV strains isolated from both the human and swine belonged to genotype IV and scattered in evolutionary branches without significant species aggregation.

CONCLUSIONIt's suggested that genotype IV HEV was the dominant genotype among the human and swine in Anqing City and probably transmitted between them in this area.

Aged ; Animals ; Base Sequence ; China ; epidemiology ; Female ; Genotype ; Hepatitis E ; epidemiology ; veterinary ; virology ; Hepatitis E virus ; classification ; genetics ; isolation & purification ; Humans ; Male ; Middle Aged ; Phylogeny ; Swine ; virology ; Swine Diseases ; epidemiology ; virology

In the present study, the genomic sequence characteristics of HN-JY40 strains of the hepatitis E virus (HEV) isolated from pigs in Henan Province, China, were analyzed and the evolutionary relationship between HN-JY40 and other sequenced strains examined. The whole genome of HN-JY40 was sequenced and analyzed by reverse transcription-polymerase chain reaction, 3' rapid amplification of cDNA ends (3' RACE) and 5' RACE. Bioinformatic analyses were carried out with Megalign, Expasy, clustal x, and MEGA 4 software. The genome of HN-JY40 was 7 223 bp in size upon removal of polyA sequences. Sizes were 9 bp and 69 bp at 5' and 3' noncoding regions, respectively. The genome of HN-JY40 was predicted to contain three open reading frames (ORFs): ORF1 (5 124 bp) encoding 1 707 amino acids; ORF2 (2 025 bp) encoding 674 amino acids; ORF3 (345 bp) encoding 114 amino acids. Phylogenetic-tree analyses indicated that HN-JY40 is a typical type-IV virus that belongs to a new subgenotype of HEV genotype 4. We sequenced and analyzed the whole genome of HN-JY40. This strategy elicited the genomic characteristics of the HEV isolated from pigs in Henan Province as well as the evolutionary relationships between HN- JY40 and other HEV isolates from pigs. We revealed that the ORF1 of HN-JY40 (153-432 nt) and human HK 104-2004 had high similarity, which offers molecular evidence for uncovering the interspecies transmission of the HEV.
Amino Acid Sequence ; Animals ; Base Sequence ; China ; Cloning, Molecular ; Genome, Viral ; Hepatitis E ; veterinary ; virology ; Hepatitis E virus ; classification ; genetics ; isolation & purification ; Humans ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; Sequence Homology, Amino Acid ; Swine ; Swine Diseases ; virology

The recent increase in the number of cases of indigenous hepatitis E virus (HEV) infection highlights the importance of identifying the transmission routes for the prevention of such infections. Presented herein is the first case of acute HEV infection after ingesting wild roe deer meat in South Korea. A 43-year-old male presented with abdominal discomfort and jaundice. He had not recently traveled abroad, but had eaten raw roe-deer meat 6-8 weeks before the presentation. On the 7th day of hospitalization the patient was diagnosed with acute viral hepatitis E. Phylogenetic analysis of his serum revealed genotype-4 HEV. This case supports the possibility of zoonotic transmission of HEV because the patient appears to have been infected with genotype-4 HEV after ingesting raw deer meat.
Adult ; Alanine Transaminase/blood ; Animals ; Bilirubin/blood ; Deer/virology ; Genotype ; Hepatitis E/*diagnosis/transmission/virology ; Hepatitis E virus/classification/*genetics/isolation & purification ; Humans ; Male ; Phylogeny ; RNA, Viral/analysis ; Republic of Korea ; Travel

The objective of current study was to investigate the quasispecies of hepatitis E virus in swine. The partial ORF2 region of HEV envelope gene from four swine HEV strains was amplified by RT-nested polymerase chain reaction (RT-nPCR). After cloning and transformation of PCR products, 20 positive clones of each HEV isolate were subject to sequencing and DNA analysis. The homology among the different clones of each isolates was 96.8%-99.7%, 98.8%-99.7%, 98.8%-99.7% and 100%, respectively, while there was 96.8%-100% sequence identity at the nucleotide level compared with HEV strains isolated in Shanghai (SAAS-JDY5). This study confirmed that there existed quasispecies of HEV in swine.
Amino Acid Sequence ; Animals ; Base Sequence ; China ; Disease Reservoirs ; virology ; Hepatitis E ; virology ; Hepatitis E virus ; chemistry ; classification ; genetics ; isolation & purification ; Mutation ; Open Reading Frames ; Phylogeny ; Sequence Alignment ; Sequence Analysis, DNA ; Swine ; virology

OBJECTIVETo explore the carrier state of hepatitis E virus(HEV) in livestock in Xi'an area.

METHODSBile samples from swine, canine, sheep and cow were collected from a local slaughtering house. Reverse transcriptase nested polymerase chain reaction (RT-nPCR) was employed to amplify the ORF2 region in HEV RNA genome. All positive samples were sequenced and compared with data from GenBank. Homology analysis was conducted based on the outcome of sequencing.

RESULTS194, 178, 79 and 191 bile samples from swine, canine, cow and sheep were collected. Positive rates with RT-nPCR method in these domestic animals were 4.10%, 0%, 0% and 0% respectively. Genetic distance analysis indicated that strains being identified were close to genotype IV of HEV, then genotype I, II and III in nucleic acid. Same outcome was shown by the same analysis on amino acid.

CONCLUSIONSwine was the only reservoir of HEV in livestock and genotype IV was the prevalent genotype.

Animals ; Animals, Domestic ; virology ; Cattle ; China ; Dogs ; Genome, Viral ; genetics ; Genotype ; Hepatitis E virus ; classification ; genetics ; isolation & purification ; Phylogeny ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sheep ; Swine

Reverse transcription-nested polymerase chain reaction (RT-nPCR) was used to detect HEV (Hepatitis E virus, HEV) RNA in dung or anus-swab samples collected from the cows with the positive anti-HEV antibodies in dairy farms in Xinjiang Autonomous Region. 7 of 60 (11.67%) cows were positive for HEV RNA in one farm. 1 of 31 (3.23%) cows was positive in the other farm. PCR amplification products were cloned, sequenced and analyzed. The result showed that the homology among the 8 bovine HEV ORF2 189bp nucleotide amplification sequences was 96.3%-100.0%, suggesting the same genotype. Compared with HEV genotype 1, 2, 3 and 4 corresponding 189 bp nucleotide sequences, the average homology was 78.5%-86.4%, 81.7%-83.8%, 79.1%-85.3% and 84.3%-95.8% respectively. The maximum homology between 8 nucleotide amplification sequences and one sequence of genotype 4 was 93.2%-95.8%. Based on the sequence of the nucleic acid fragments, a phylogenetic tree was constructed. It was illustrated that 8 bovine HEV ORF2 189bp nucleotide amplification sequences in this study and human C5 strain, swine swC3, swXJ strain belonged to genotype 4. The finding suggested that infection of HEV probably existed in the cow group in Xinjiang Autonomous Region and the cow might be a new animal host except swine in origin of HEV infection.
Animals ; Cattle ; virology ; China ; Disease Reservoirs ; virology ; Feces ; virology ; Hepatitis Antibodies ; blood ; Hepatitis E ; immunology ; virology ; Hepatitis E virus ; classification ; genetics ; immunology ; isolation & purification ; Humans ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; RNA, Viral ; genetics ; isolation & purification ; Sequence Analysis, DNA ; Swine ; virology