title: Detecting hepatitis E virus with a reverse transcription polymerase chain reaction enzyme-linked immunosorbent assay.
abstract: This study aimed to develop a specific and sensitive reverse transcription polymerase chain reaction enzyme-linked immunosorbent assay (RT-PCR-ELISA) for detecting hepatitis E virus (HEV). Eight sets of primers and biotinylated probes designed in the ORF2-ORF3 overlapping region of HEV were tested for sensitivity. The ability of nested reverse transcription polymerase chain reaction (RT-PCR) and RT-PCR-ELISA to detect HEV was compared. RT-PCR-ELISA was 10-100 times more sensitive than nested RT-PCR and could detect 0.01 ng/μl HEV in swine stool samples. In terms of specificity, RT-PCR-ELISA did not falsely detect HEV when other viruses such as hepatitis A virus, rotavirus, norovirus genotype I, norovirus genotype II, and Feline calicivirus were present. Therefore, RT-PCR-ELISA appears to be a sensitive and specific method for detecting HEV.
authors: Department of Food and Nutrition, School of Food Science and Technology, Chung-Ang University, 72-1, Nae-ri, Daeduck-myun, Ansung-si, Kyounggi-do, 456-756, South Korea.
authors: Dong Joo
authors: DJ
authors: Seo
authors: Hongmin
authors: H
authors: Tahk
authors: Kang Bum
authors: KB
authors: Lee
authors: Min Hwa
authors: MH
authors: Lee
authors: Na Ry
authors: NR
authors: Son
authors: Sheungwoo
authors: S
authors: Seo
authors: Doo-Sung
authors: DS
authors: Cheon
authors: Bog-Hieu
authors: BH
authors: Lee
authors: Changsun
authors: C
authors: Choi
mesh_terms: D000818:Animals; D019921:Biotinylation; D017931:DNA Primers / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D005243:Feces / Q000821:virology; D016751:Hepatitis E / Q000821:virology*; D016752:Hepatitis E virus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000821:virology*
chemical_list: D017931:DNA Primers; D012367:RNA, Viral