title: Optimization of the elution buffer and concentration method for detecting hepatitis E virus in swine liver using a nested reverse transcription-polymerase chain reaction and real-time reverse transcription-polymerase chain reaction.
abstract: The aim of this study was to develop an optimal technique for detecting hepatitis E virus (HEV) in swine livers. Here, three elution buffers and two concentration methods were compared with respect to enhancing recovery of HEV from swine liver samples. Real-time reverse transcription-polymerase chain reaction (RT-PCR) and nested RT-PCR were performed to detect HEV RNA. When phosphate-buffered saline (PBS, pH 7.4) was used to concentrate HEV in swine liver samples using ultrafiltration, real-time RT-PCR detected HEV in 6 of the 26 samples. When threonine buffer was used to concentrate HEV using polyethylene glycol (PEG) precipitation and ultrafiltration, real-time RT-PCR detected HEV in 1 and 3 of the 26 samples, respectively. When glycine buffer was used to concentrate HEV using ultrafiltration and PEG precipitation, real-time RT-PCR detected HEV in 1 and 3 samples of the 26 samples, respectively. When nested RT-PCR was used to detect HEV, all samples tested negative regardless of the type of elution buffer or concentration method used. Therefore, the combination of real-time RT-PCR and ultrafiltration with PBS buffer was the most sensitive and reliable method for detecting HEV in swine livers.
authors.affiliation: Department of Food and Nutrition, Chung-Ang University, Republic of Korea.
authors.forename: Na Ry
authors.initials: NR
authors.lastname: Son
authors.affiliation: Department of Food and Nutrition, Chung-Ang University, Republic of Korea.
authors.forename: Dong Joo
authors.initials: DJ
authors.lastname: Seo
authors.affiliation: Department of Food and Nutrition, Chung-Ang University, Republic of Korea.
authors.forename: Min Hwa
authors.initials: MH
authors.lastname: Lee
authors.affiliation: Department of Food and Nutrition, Chung-Ang University, Republic of Korea.
authors.forename: Sheungwoo
authors.initials: S
authors.lastname: Seo
authors.affiliation: Department of Food and Nutrition, Chung-Ang University, Republic of Korea.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Food and Nutrition, Chung-Ang University, Republic of Korea; School of Food Science and Technology, Chung-Ang University, Republic of Korea.
authors.forename: Bog-Hieu
authors.initials: BH
authors.lastname: Lee
authors.affiliation: Food Microbiology Division, Ministry of Food and Drug Safety, Republic of Korea.
authors.forename: Jeong-Su
authors.initials: JS
authors.lastname: Lee
authors.affiliation: Food Microbiology Division, Ministry of Food and Drug Safety, Republic of Korea.
authors.forename: In-Sun
authors.initials: IS
authors.lastname: Joo
authors.affiliation: Food Microbiology Division, Ministry of Food and Drug Safety, Republic of Korea.
authors.forename: In-Gyun
authors.initials: IG
authors.lastname: Hwang
authors.affiliation: Department of Food and Nutrition, Chung-Ang University, Republic of Korea; School of Food Science and Technology, Chung-Ang University, Republic of Korea. Electronic address: cchoi@cau.ac.kr.
authors.forename: Changsun
authors.initials: C
authors.lastname: Choi
mesh_terms: D000818:Animals; D002021:Buffers; D011232:Chemical Precipitation; D005374:Filtration / Q000379:methods; D016751:Hepatitis E / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D016752:Hepatitis E virus / Q000235:genetics / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology; D013997:Time Factors
chemical_list: D002021:Buffers
keywords: Concentration; Elution; Hepatitis E virus (HEV); Nested reverse transcription-polymerase chain reaction (nested RT-PCR); Real-time reverse transcription-polymerase chain reaction (real-time RT-PCR)