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2015 | 18 | 4 |

Tytuł artykułu

A multiplex PCR for simultaneous detection of classical swine fever virus, African swine fever virus, highly pathogenic porcine reproductive and respiratory syndrome virus, porcine reproductive and respiratory syndrome virus and pseudorabies in swines

Treść / Zawartość

Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
In this assay, we developed and evaluated a multiplex PCR (mPCR) for its ability in detecting multiple infections of swine simultaneously. Four pairs of primers were used to detect five viruses. Specific primers were designed for classical swine fever virus (CSFV), African swine fever virus (ASFV) and pseudorabies (PRV). A pair of primers was designed prudently for two different types of porcine reproductive and respiratory syndrome virus that respectively were porcine reproductive and respiratory syndrome virus (PRRSV), highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV). The detection limits of the mPCR were 1.09×10⁴, 1.50×10³, 2.10×10³, 1.30×10³ and 8.97×10² copies/reaction for CSFV, ASFV, HP-PRRSV, PRRSV and PRV, respectively. A total of 49 clinical specimens were tested by the mPCR, and the result showed that co-infection by two or three viruses was 51%. In conclusion, the PCR is a useful tool for clinical diagnosis of not only single infections but also mixed infections in swines.

Słowa kluczowe

Wydawca

-

Rocznik

Tom

18

Numer

4

Opis fizyczny

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Twórcy

autor
  • Animal Quarantine Laboratory,College of Veterinary Medicine, Sichuan Agricultural University, Chengdu Sichuan 611130, China
autor
  • Animal Quarantine Laboratory,College of Veterinary Medicine, Sichuan Agricultural University, Chengdu Sichuan 611130, China
autor
  • Animal Quarantine Laboratory,College of Veterinary Medicine, Sichuan Agricultural University, Chengdu Sichuan 611130, China
autor
  • Animal Quarantine Laboratory,College of Veterinary Medicine, Sichuan Agricultural University, Chengdu Sichuan 611130, China
autor
  • Animal Quarantine Laboratory,College of Veterinary Medicine, Sichuan Agricultural University, Chengdu Sichuan 611130, China
autor
  • Animal Quarantine Laboratory,College of Veterinary Medicine, Sichuan Agricultural University, Chengdu Sichuan 611130, China
autor
  • Animal Quarantine Laboratory,College of Veterinary Medicine, Sichuan Agricultural University, Chengdu Sichuan 611130, China
  • Key laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu Sichuan 611130, China

Bibliografia

  • Aguero M, Fernhndez J, Romero L, Zamora MJ, Shnchez C, Belhk S, Arias M, Shnchez-Vizcatno JM (2004) A highly sensitive and specific gel-based multiplex RT-PCR assay for the simultaneous and differential diagnosis of African swine fever and classical swine fever in clinical samples. Vet Res 35: 551-563.
  • Cao S, Chen H, Zhao J, Lü J, Xiao S, Jin M, Guo A, Wu B, He Q (2005) Detection of porcine circovirus type 2, porcine parvovirus and porcine pseudorabies virus from pigs with postweaning multisystemic wasting syndrome by multiplex PCR. Vet Res Commun 29: 263-269.
  • Dong XT, Li YF, Jiang P, Xu JR (2006) Detection of co-infections of porcine reproductive and respiratory syndrome virusand classical swine fever virus. Animal Husbandry & Vet Med 5: 006.
  • Durzyńska J, Pacholska-Bogalska J, Kaczmarek M, Hanć T, Durda M, Skrzypczak M, Goździcka-Józefiak A (2011) Multiplex PCR for identification of herpes virus infections in adolescents. J Med Virol 83: 267-271.
  • Elnifro EM, Ashshi AM, Cooper RJ, Klapper PE (2000) Multiplex PCR: optimization and application in diagnostic virology. Clin Microbiol Rev 13: 559-570.
  • Gallardo C, Ademun AR, Nieto R, Nantima N, Arias M, Marttn E, Pelayo V, Bishop RP (2013) Genotyping of African swine fever virus (ASFV) isolates associated with disease outbreaks in Uganda in 2007. Afr J Biotechnol 10: 3488-3497.
  • Giammarioli M, Pellegrini C, Casciari C, De Mia GM (2008) Development of a novel hot-start multiplex PCR for simultaneous detection of classical swine fever virus, African swine fever virus, porcine circovirus type 2, porcine reproductive and respiratory syndrome virus and porcine parvovirus.Vet Res Commun 32: 255-262.
  • Giridharan P, Hemadri D, Tosh C, Sanyal A, Bandyopadhyay SK (2005) Development and evaluation of a multiplex PCR for differentiation of foot-and-mouth disease virus strains native to India. J Virol Methods 126: 1-11.
  • Jiang Y, Shang H, Xu H, Zhu L, Chen W, Zhao L, Fang L (2010) Simultaneous detection of porcine circovirus type 2, classical swine fever virus, porcine parvovirus and porcine reproductive and respiratory syndrome virus in pigs by multiplex polymerase chain reaction. Vet J 183: 172-175.
  • King DP, Reid SM, Hutchings G.H, Grierson SS, Wilkinson PJ, Dixon LK., Bastos AD, Drew T W (2003) Development of a TaqMan PCR assay with internal amplification control for the detection of African swine fever virus. J Virol Methods 107: 53-61.
  • Lee CS, Moon HJ, Yang JS, Park SJ, Song DS, Kang BK., Park BK. (2007) Multiplex PCR for the simultaneous detection of pseudorabies virus, porcine cytomegalovirus, and porcine circovirus in pigs. J Virol Methods 139: 39-43.
  • Liu S, Zhao Y, Hu Q, Lv C, Zhang C, Zhao R, Hu F, Lin W, Cui S (2011) A multiplex RT-PCR for rapid and simultaneous detection of porcine teschovirus, classical swine fever virus, and porcine reproductive and respiratory syndrome virus in clinical specimens. J Virol Methods 172: 88-92.
  • Ogawa H, Taira O, Hirai T, Takeuchi H, Nagao A, Ishikawa Y, Tuchiya K., Nunoya T, Ueda S, (2009) Multiplex PCR and multiplex RT-PCR for inclusive detection of major swine DNA and RNA viruses in pigs with multiple infections. J Virol Methods 160: 210-214.
  • Penrith ML (2009) African swine fever. Onderstepoort J Vet Res 76: 91-95.
  • Wernike K., Hoffmann B, Beer M (2013).Single-tube multiplexed molecular detection of endemic porcine viruses in combination with background screening for transboundary diseases. J Clin Microbiol 51: 938-944.
  • Yu X, Chen N, Wang L, Wu J, Zhou Z, Ni J, Li X, Zhai X, Shi J, Tian K (2012) New genomic characteristics of highly pathogenic porcine reproductive and respiratory syndrome viruses do not lead to significant changes in pathogenicity. Vet Microbiol 158: 291-299.
  • Yue F, Cui S, Zhang C, Yoon KJ (2009) A multiplex PCR for rapid and simultaneous detection of porcine circovirus type 2, porcine parvovirus, porcine pseudorabies virus, and porcine reproductive and respiratory syndrome virus in clinical specimens.Virus genes 38: 392-397.
  • Zanella EL, Miller LC, Lager KM, Bigelow TT (2012) Evaluation of a real-time polymerase chain reaction assay for Pseudorabies virus surveillance purposes. J Vet Diagn Invest 24: 739-745.
  • Zhao JJ, Cheng D, Li N, Sun Y, Shi Z, Zhu Q H, Tu C, Tong G.Z, Qiu HJ (2008) Evaluation of a multiplex real-time RT-PCR for quantitative and differential detection of wild-type viruses and C- train vaccine of Classical swine fever virus.Vet Microbiol 126: 1-10.
  • Zhao YZ, He DS, Zhang YP, Su DP (2010) Establishment of a Multiplex PCR Assay for Simultaneous Detection of PCV2, PPV and PRV.China Animal Husbandry & Veterinary Medicine 2: 10-21.
  • Zhou L, Yang H (2010) Porcine reproductive and respiratory syndrome in China.Virus Res 154: 31-37.
  • Zhou Z, Li X, Liu Q, Hu D, Yue X., Ni J, Yu X, Zhai X, Chen N (2012) Complete genome sequence of two novel Chinese virulent porcine reproductive and respiratory syndrome virus variants. J Virol 86: 6373-6374.
  • Zhou Z, Ni J, Cao Z, Han X, Xia Y, Zi Z, Ning K, Liu Q, Cai L, Qiu P (2011) The epidemic status and genetic diversity of 14 highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) isolates from China in 2009. Vet Microbiol 150: 257-269.

Typ dokumentu

Bibliografia

Identyfikatory

Identyfikator YADDA

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