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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
immunology and microbiology
Multiplex nested RT-PCR for the detection of porcine enteric viruses
Journal of Virological Methods, Volume 165, No. 2, Year 2010
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Description
Porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), and porcine group A rotavirus (PRV-A) are major viruses causing enteric diseases of piglets. A multiplex nested reverse transcription polymerase chain reaction (multiplex nested RT-PCR) was developed for the detection of these viruses in field samples from piglets with diarrhea. A mixture of (1) three external pairs of primers, yielding in the amplification step two different amplicons with sizes of 950bp and 317bp and (2) three pairs of internal primers in a second round of PCR (nested PCR), yielding two different amplicons with sizes of 792bp and 208bp for TGEV and porcine PRV-A, respectively. The genome of PEDV was not detected after the amplification step but it was detected in the second round of PCR, yielding amplicon with size of 291bp. Multiplex nested RT-PCR can detect TGEV, PRV-A, and PEDV up to concentration 102TCID50/mL, 101TCID50/mL, and 27.2μg/μl of RNA, respectively. A total of 175 field samples were collected from swine with diarrhea from January 2005 until July 2007. The samples were tested for the presence of three viruses by a multiplex nested RT-PCR. Dual infections with PEDV and PRV-A were identified in seven specimens (4%) (n=6). Twenty-one (25%) infections were caused by PEDV and thirty-four infections (41%) were caused by PRV-A. The genome of TGEV was not detected in any of these field samples, however TGEV was detected in piglets infected experimentally. The multiplex nested RT-PCR is rapid, sensitive, and a cost-effective detection method for the detection of porcine enteric viruses. © 2010 Elsevier B.V.
Authors & Co-Authors
Abid, Nabil Ben Salem
Russian Federation, Vladimir
Federal State-financed Institution Federal Centre for Animal Health
Tunisia, Monastir
Faculté de Pharmacie de Monastir
Chupin, Sergei A.
Russian Federation, Vladimir
Federal State-financed Institution Federal Centre for Animal Health
Byadovskaya, Olga P.
Russian Federation, Vladimir
Federal State-financed Institution Federal Centre for Animal Health
Andreeva, Olga G.
Russian Federation, Vladimir
Federal State-financed Institution Federal Centre for Animal Health
Aouni, Mahjoub
Tunisia, Monastir
Faculté de Pharmacie de Monastir
Prokhvatilova, Larisa B.
Russian Federation, Vladimir
Federal State-financed Institution Federal Centre for Animal Health
Statistics
Citations: 37
Authors: 6
Affiliations: 2
Identifiers
Doi:
10.1016/j.jviromet.2010.02.010
ISSN:
01660934