Detection of transmissible gastroenteritis virus by RT-PCR and differentiation from porcine respiratory coronavirus

• Published in: Journal of virological methods Vol. 66; no. 2; pp. 303 - 309
• Main Authors: Paton, David, Ibata, Georgina, Sands, Jenny, McGoldrick, Adrian
• Format: Journal Article
• Language: English
• Published: London Elsevier B.V 01.07.1997; Amsterdam Elsevier; New York, NY Published by Elsevier B.V
• Subjects: Animals; Biological and medical sciences; Coronavirus - genetics; Coronavirus - isolation & purification; Coronavirus Infections - diagnosis; Coronavirus Infections - veterinary; Diagnosis, Differential; Feces - virology; Fundamental and applied biological sciences. Psychology; Gastroenteritis virus; Gastroenteritis, Transmissible, of Swine - diagnosis; Intestine, Small - virology; Membrane Glycoproteins - genetics; Microbiology; Molecular Sequence Data; Polymerase Chain Reaction - methods; Respiratory coronavirus; RNA, Viral - analysis; Sensitivity and Specificity; Serial Passage; Spike Glycoprotein, Coronavirus; Swine; Techniques used in virology; Transmissible gastroenteritis; Transmissible gastroenteritis virus - genetics; Transmissible gastroenteritis virus - isolation & purification; Viral Envelope Proteins - genetics; Virology; Gastroenteritis virus; Respiratory coronavirus; Transmissible gastroenteritis; RNA; Reverse transcription; Method; Porcine transmissible gastroenteritis virus; Virus; Polymerase chain reaction; Vertebrata; Coronavirus; Mammalia; Swine; Artiodactyla; Coronaviridae; Veterinary; Detection; Porcine respiratory coronavirus; Ungulata
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/S0166-0934(97)00055-4

An RT-PCR method was developed that amplified genetic material from the 5′ end of the S protein gene of both transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV), but discriminated between the two by the size of the product generated. A number of restriction endonuclease enzymes were assessed for recognition of the amplicons so produced. The assay was shown to detect viral RNA from all of the 26 different TGEV and PRCV isolates examined, covering a period from 1946 to 1996. Detection of TGEV in clinical specimens was possible using a spin column method to extract RNA and sensitivity was compared to virus isolation and antigen detection ELISA. The method could provide a means of confirming positive results from immunological screening tests such as FAT and ELISA, reducing the need for virus isolation and convalescent serology.