Detection of transmissible gastroenteritis virus by RT-PCR and differentiation from porcine respiratory coronavirus.
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.
Paton
D
D
Virology Department, Central Veterinary Laboratory (Weybridge), Surrey, UK. dpaton.wood.cv1@gtnet.gov.uk
Ibata
G
G
Sands
J
J
McGoldrick
A
A
D000818:Animals; D017934:Coronavirus / Q000235:genetics / Q000302:isolation & purification*; D018352:Coronavirus Infections / Q000175:diagnosis / Q000662:veterinary*; D003937:Diagnosis, Differential; D005243:Feces / Q000821:virology; D005761:Gastroenteritis, Transmissible, of Swine / Q000175:diagnosis*; D007421:Intestine, Small / Q000821:virology; D008562:Membrane Glycoproteins / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis; D012680:Sensitivity and Specificity; D012692:Serial Passage; D064370:Spike Glycoprotein, Coronavirus; D013552:Swine; D005760:Transmissible gastroenteritis virus / Q000235:genetics / Q000302:isolation & purification*; D014759:Viral Envelope Proteins / Q000235:genetics
D008562:Membrane Glycoproteins; D012367:RNA, Viral; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins; C578557:spike glycoprotein, SARS-CoV; C578553:spike protein, mouse hepatitis virus