Differential detection of turkey coronavirus, infectious bronchitis virus, and bovine coronavirus by a multiplex polymerase chain reaction

The objective of the present study was to develop a multiplex polymerase chain reaction (PCR) method for differential detection of turkey coronavirus (TCoV), infectious bronchitis coronavirus (IBV), and bovine coronavirus (BCoV). Primers were designed from conserved or variable regions of nucleocaps...

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Bibliographic Details
Published in:	Journal of virological methods Vol. 131; no. 1; pp. 86 - 91
Main Authors:	Loa, C.C., Lin, T.L., Wu, C.C., Bryan, T.A., Hooper, T.A., Schrader, D.L.
Format:	Journal Article
Language:	English
Published:	London Elsevier B.V 01-01-2006 Amsterdam Elsevier New York, NY
Subjects:	Animals Astrovirus Biological and medical sciences Bovine Bovine coronavirus Coronavirus Coronavirus Infections - diagnosis Coronavirus, Bovine - chemistry Coronavirus, Bovine - genetics Coronavirus, Turkey - chemistry Coronavirus, Turkey - genetics DNA Primers Enterovirus Escherichia coli Fundamental and applied biological sciences. Psychology Genes, Viral Infectious bronchitis virus Infectious bronchitis virus - chemistry Infectious bronchitis virus - genetics Infectious bursal disease virus Marek's disease herpesvirus Membrane Glycoproteins - genetics Microbiology Multiplex Mycoplasma gallisepticum Newcastle disease virus Nucleocapsid Proteins - genetics Polymerase chain reaction Polymerase Chain Reaction - methods Reovirus RNA, Viral - genetics Salmonella enterica Sensitivity and Specificity Species Specificity Spike Glycoprotein, Coronavirus Techniques used in virology Turkey Turkey coronavirus Turkeys Viral Envelope Proteins - genetics Virology Polymerase chain reaction Turkey Coronavirus Bovine Multiplex Infectious bronchitis virus Bovine coronavirus Microbiology Multiplex polymerase chain reaction Avian infectious bronchitis virus Method Virology Virus Vertebrata Mammalia Turkey coronavirus Artiodactyla Coronaviridae Veterinary Detection Nidovirales Ungulata
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Summary:	The objective of the present study was to develop a multiplex polymerase chain reaction (PCR) method for differential detection of turkey coronavirus (TCoV), infectious bronchitis coronavirus (IBV), and bovine coronavirus (BCoV). Primers were designed from conserved or variable regions of nucleocapsid (N) or spike (S) protein gene among TCoV, IBV, and BCoV and used in the same PCR reaction. Reverse transcription followed by the PCR reaction was used to amplify a portion of N or S gene of the corresponding coronaviruses. The PCR products were detected on agarose gel stained with ethidium bromide. Two PCR products, a 356-bp band corresponding to N gene and a 727-bp band corresponding to S gene, were obtained for TCoV isolates. In contrast, one PCR product of 356 bp corresponding to a fragment of N gene was obtained for IBV strains and one PCR product of 568 bp corresponding to a fragment of S gene was obtained for BCoV. There were no PCR products with the same primers for Newcastle disease virus, Marek's disease virus, turkey pox virus, pigeon pox virus, fowl pox virus, reovirus, infectious bursal disease virus, enterovirus, astrovirus, Salmonella enterica, Escherichia coli, and Mycoplasma gallisepticum. Performance of the assay with serially diluted RNA demonstrated that the multiplex PCR could detect 4.8 × 10 −3 μg of TCoV RNA, 4.6 × 10 −4 μg of IBV RNA, and 8.0 × 10 −2 μg of BCoV RNA. These results indicated that the multiplex PCR as established in the present study is a rapid, sensitive, and specific method for differential detection of TCoV, IBV, and BCoV in a single PCR reaction.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0166-0934 1879-0984
DOI:	10.1016/j.jviromet.2005.07.007