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KMID : 0893420170180020253
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Journal of Veterinary Science
2017 Volume.18 No. 2 p.253 ~ p.256
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Duplex nested reverse transcriptase polymerase chain reaction for simultaneous detection of type 2 porcine reproductive and respiratory syndrome virus and porcine circovirus type 2 from tissue samples
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Kim Hye-Kwon
Lyoo Kwang-Soo
Huynh Thi My Le
Moon Hyoung-Joon
Nguyen Van Giap
Park Bong-Kyun
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Abstract
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There are high levels of co-incidence of porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) in porcine tissue. This study established a duplex nested reverse transcriptase polymerase chain reaction (RT-PCR) method that targets the genomic RNA of type 2 PRRSV and the mRNA of PCV2 in infected tissues. The method amplified discriminative bands of 347 bp and 265 bp specific for type 2 PRRSV and PCV2, respectively. The limits of detection of the duplex nested RT-PCR were 101.5 TCID50/mL for type 2 PRRSV and 102 infected cells/mL for PCV2. The kappa statistic, which measures agreement between methods, was 0.867, indicating a good level of agreement. This RNA-based duplex RT-PCR approach can be another way to detect type 2 PRRSV and PCV2 simultaneously and with improved convenience.
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KEYWORD
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duplex nested RT-PCR, porcine circovirus type 2, type 2 porcine reproductive and respiratory syndrome virus
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