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Aanalysis of Genotype and ORF3 Gene Mutation of Hepatitis E Virus in Wuhan

Author: ZhuYongBao
Tutor: HuangYuanCheng
School: Huazhong University of Science and Technology
Course: Internal Medicine
Keywords: Hepatitis virus, HEV Popular features Genotype Subtypes Open reading frame Gene mutation
CLC: R512.6
Type: Master's thesis
Year: 2011
Downloads: 8
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Background Hepatitis E is caused by the hepatitis E virus (hepatitis E virus, HEV) an acute self-limiting disease, mainly by the fecal - oral transmission, sporadic outbreaks in two forms. Patients with clinical manifestations similar to hepatitis A, but mortality is high, especially compared to the other hepatitis infection in pregnant women up to 30% to 100% mortality. China is one of the areas of high incidence of hepatitis E, Xinjiang Uygur Autonomous Region in the 1986 to 1988 period occurred far the world's largest hepatitis E epidemic, a total of 119,280 cases, 707 people were killed, to the community and the economy has caused a great deal of harm. In recent years, the incidence of acute sporadic hepatitis E has continued to rise. HEV is a single-stranded positive RNA virus, non-enveloped, the genome of about 7.3 kb, containing the three open reading frames (ORFs), wherein ORF1 is located in the genome of the 5 'end of the coding RNA replication of the non-structural proteins , ORF2 is located in the genomic 3 'end encoding a viral capsid protein, ORF3 also located in the genomic 3' end encodes a phosphoprotein, and its function has not yet been fully elucidated. Ⅰ, Ⅱ and Ⅲ the HEV ORF3 gene fragment length 372 bp, encoding a protein of 123 amino acids, and the gene type IV HEV insert in its ORF3 nucleotide (U), resulting in a reading frame shift, ORF3 encoding the compared with the other genotypes, the protein N-terminal deletion of nine amino acids, containing 114 amino acids. The subject of general Wuhan Tongji Hospital infection outpatient and inpatient department with acute hepatitis E patients (such as gender and age) investigation and HEV genotyping, and gene type Ⅳ HEV ORF3 gene and its encoded protein variants situation detection and analysis. Objective 1. Explore Wuhan area popular features of sporadic acute hepatitis E virus genotype clear the Wuhan region HEV genotype distribution provides some clues. 2. Study the Wuhan region gene HEV IV of ORF3 gene and its encoded protein variation, in order to further explore the clinical significance of the genetic type Ⅳ the HEV ORF3 gene mutation lay the foundation. Method 1. Collected in our hospital from April 2009 to June 2010, 162 infected with outpatient and inpatient department with acute hepatitis E serum stored at -80 ℃ to be tested, after dispensing. Investigate the general condition of patients (such as gender and age). 2 RT - nested polymerase chain reaction (RT-nPCR) detected HEV RNA serum of 162 patients with acute hepatitis, and PCR products were sequenced, and sequencing results HEV homologous evolution analysis. 3. Ⅳ type 41 gene HEV RNA gene fragments (5020 ~ 5392nt and 5347 to 5956nt, EF570133) was amplified by reverse transcription - nested polymerase chain reaction (RT-nPCR), and PCR products were sequenced , then use ContigExpress sequencing results stitching (containing HEV ORF3 sequence), MegAlign (DNAStar package) to compare the HEV ORF3, view the the HEV ORF3 gene and its encoded protein variation. 1 male to female ratio of the Wuhan regional acute hepatitis patients 4.40:1,30 to 59-year-old age group accounted for 71.60% of 162 patients with acute hepatitis serum, 41 serum HEV RNA positive rate was 25.31% homologous evolutionary analysis showed that 41 HEV are genetically type Ⅳ, 84.7% to 100% nucleotide homology between the strains, gene type Ⅳ can be further divided into five subtypes, namely Ⅳ-A , IV-B, IV-C, IV-D of Ⅳ-E subtype within each subtype nucleotide homology of 91.3% to 100%, 98.0%, 92.0% and 100%, 98.0% ~ 100%, 100%, Ⅳ-A subtypes and Guangxi strains Shanghai the strains and Xi'an strains, nucleotide homology were 91.3% to 96.0%, from 92.0% to 97.3% and 94.7% to 99.3%, Ⅳ - the subtype D Beijing strain nucleotide homology of 95.3% to 96.7%. 2.41 gene type Ⅳ HEV RNA gene fragments were amplified from strains 18, 18 HEV ORF3 a total of 47 sites detected the presence of nucleotide substitutions, including 21 replacement occurred in the first codon one, two nucleotides, 26 replacement occurs in the third codon nucleotide; its encoded protein, there are 19 sites of amino acid variation. Conclusion. Wuhan acute hepatitis E is more common in men, good hair in the 30 to 59 age group, and are caused by gene type IV HEV gene type Ⅳ can be further divided into IV-A, IV-B, IV - C, Ⅳ-D and Ⅳ-E 5 subtypes, Ⅳ-A subtypes and strains Guangxi, Shanghai strains and Xi'an strains closer genetic relationship, IV-D subtypes closer kinship with Beijing strains. Wuhan region gene type Ⅳ HEV ORF3 nucleotide substitution, there are several sites, these sites variation in acute hepatitis progression, what is the significance needs further study.

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CLC: > Medicine, health > Internal Medicine > Infectious disease > Viral infections > Viral Hepatitis
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