title: The survey of porcine teschoviruses in field samples in China with a universal rapid probe real-time RT-PCR assay.
abstract: A real-time reverse transcription polymerase chain reaction (RT-PCR) based on TaqMan was established and evaluated for quantitative detection of porcine teschoviruses (PTVs). A pair of primers and a TaqMan probe targeting on the highly conserved sequence of the 5'-untranslated region (5'-UTR) of one to 11 serotypes of PTV were designed. Standard plasmid DNA containing PCR amplification of the 5'-UTR were constructed and used to develop the real-time RT-PCR. The results indicated that the real-time RT-PCR was specific for detection of PTV with a detection limit of 10 copies/μL, but not for porcine parvovirus, porcine circovirus type 2, porcine reproductive and respiratory syndrome virus, pseudorabies virus, classical swine fever virus. The coefficient of variation of inter-assay and intra-assay were less than 3 %. A total of 91 clinical samples were tested by the real-time RT-PCR and virus isolation (OIE 2008) and positive rates were 79.12 % (72/91) and 57.14 % (48/91), respectively. In conclusion, the developed real-time RT-PCR assay was an effective method for detection and quantification of PTV in fields or organs of infected pigs.
authors.affiliation: Department of Preventive, VeterinaryCollege of Veterinary Medicine, China Agricultural University, Beijing, 100094, China.
authors.forename: Chaofan
authors.initials: C
authors.lastname: Zhang
authors.forename: Zhongtian
authors.initials: Z
authors.lastname: Wang
authors.forename: Feng
authors.initials: F
authors.lastname: Hu
authors.forename: Yebing
authors.initials: Y
authors.lastname: Liu
authors.forename: Zheng
authors.initials: Z
authors.lastname: Qiu
authors.forename: Shun
authors.initials: S
authors.lastname: Zhou
authors.forename: Shangjin
authors.initials: S
authors.lastname: Cui
authors.forename: Ming
authors.initials: M
authors.lastname: Wang
mesh_terms: D020121:5' Untranslated Regions; D000818:Animals; D002681:China / Q000453:epidemiology; D057230:Limit of Detection; D010850:Picornaviridae Infections / Q000175:diagnosis / Q000453:epidemiology / Q000662:veterinary* / Q000821:virology; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods / Q000592:standards / Q000662:veterinary*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology*; D052700:Teschovirus / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D020121:5' Untranslated Regions; D012367:RNA, Viral