Multiplex PCR and multiplex RT-PCR for inclusive detection of major swine DNA and RNA viruses in pigs with multiple infections

被引:69
作者
Ogawa, Hirohito [1 ]
Taira, Osamu [1 ]
Hirai, Takuya [2 ]
Takeuchi, Hiromi [1 ]
Nagao, Aki [1 ]
Ishikawa, Yoshiki [1 ]
Tuchiya, Kotaro [1 ]
Nunoya, Tetsuo [1 ]
Ueda, Susumu [1 ]
机构
[1] Nippon Inst Biol Sci, Tokyo 1980024, Japan
[2] Miyazaki Univ, Dept Vet Pathol, Fac Agr, Miyazaki 8892192, Japan
关键词
Swine viruses; Porcine circovirus type 2; Multiplex PCR; Multiplex RT-PCR; MULTISYSTEMIC WASTING SYNDROME; TRANSMISSIBLE GASTROENTERITIS VIRUS; EPIDEMIC DIARRHEA VIRUS; ISOLATE ATCC VR-2332; PORCINE CIRCOVIRUS; DIFFERENTIAL DETECTION; DISEASE; PRRSV; COINFECTION; INFERTILITY;
D O I
10.1016/j.jviromet.2009.05.010
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Multiplex PCR and multiplex RT-PCR were developed to identify nine viruses in pigs with multiple infections. These viruses are: porcine circovirus type 2 (PCV2), suid herpesvirus 1, porcine parvovirus (PPV), porcine reproductive and respiratory syndrome virus (PRRSV), Japanese encephalitis virus, porcine rotavirus A (PoRV-A), porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), and Getah virus. These methods were shown to be high specificity and sensitivity. In the clinical application, a total of 75 field samples were examined by our methods and previously reported methods for PCV2, PRRSV, TGEV, and PEDV. As a result, the detection rates of our multiplex PCR and multiplex RTPCR were higher than those of the previously reported methods. Furthermore, it was confirmed that 24 PCV2 positive samples were co-infected with other viruses, 11 with PRRSV, 10 with PPV, 2 with PoRV-A, and 1 with TGEV by a combination of multiplex PCR and multiplex RT-PCR. PPV and PoRV-A were newly detected by multiplex PCR and multiplex RT-PCR. These results suggest that the combination of our multiplex PCR and multiplex RT-PCR is useful for rapid and accurate identification of nine major pathogenic viruses in pigs with multiple infections. (C) 2009 Elsevier B.V. All rights reserved.
引用
收藏
页码:210 / 214
页数:5
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