The survey of porcine teschoviruses in field samples in China with a universal rapid probe real-time RT-PCR assay
Identifieur interne : 002304 ( Main/Exploration ); précédent : 002303; suivant : 002305The survey of porcine teschoviruses in field samples in China with a universal rapid probe real-time RT-PCR assay
Auteurs : Chaofan Zhang [République populaire de Chine] ; Zhongtian Wang [République populaire de Chine] ; Feng Hu [République populaire de Chine] ; Yebing Liu [République populaire de Chine] ; Zheng Qiu [République populaire de Chine] ; Shun Zhou [République populaire de Chine] ; Shangjin Cui [République populaire de Chine] ; Ming Wang [République populaire de Chine]Source :
- Tropical Animal Health and Production [ 0049-4747 ] ; 2013-04-01.
English descriptors
Abstract
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.
Url:
DOI: 10.1007/s11250-012-0312-0
Affiliations:
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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. 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