Development of a reverse transcription-nested polymerase chain reaction assay for differential diagnosis of transmissible gastroenteritis virus and porcine respiratory coronavirus from feces and nasal swabs of infected pigs

Citation
L. Kim et al., Development of a reverse transcription-nested polymerase chain reaction assay for differential diagnosis of transmissible gastroenteritis virus and porcine respiratory coronavirus from feces and nasal swabs of infected pigs, J VET D INV, 12(4), 2000, pp. 385-388
Citations number
13
Categorie Soggetti
Veterinary Medicine/Animal Health
Journal title
JOURNAL OF VETERINARY DIAGNOSTIC INVESTIGATION
ISSN journal
10406387 → ACNP
Volume
12
Issue
4
Year of publication
2000
Pages
385 - 388
Database
ISI
SICI code
1040-6387(200007)12:4<385:DOARTP>2.0.ZU;2-Y
Abstract
Transmissible gastroenteritis virus (TGEV), a coronavirus, replicates in in testinal enterocytes and causes diarrhea in young pigs. Porcine respiratory coronavirus (PRCV), a spike (S) gene natural deletion mutant of TGEV, has a respiratory tissue tropism and causes mild or subclinical respiratory inf ections. Conventional antigen-based diagnostic tests fail to differentiate TGEV and PRCV, and a blocking ELISA test to serologically differentiate TGE V/PRCV-infected pigs is conducted on convalescent serum retrospectively aft er disease outbreaks. A reverse transcription (RT)-nested polymerase chain reaction (PCR) with primers targeted to the S gene deletion region to diffe rentiate TGEV/PRCV was developed. The specificity of the RT-nested PCR was confirmed with reference and recent field strains of TGEV/PRCV, and its sen sitivity was analyzed by testing nasal and fecal samples collected from pig s at various days postinoculation (DPI) with TGEV or PRCV. Specific PCR pro ducts for TGEV/PRCV were detected only with the homologous reference or fie ld coronaviruses and for 10-14 DPI of pigs with TGEV (feces) or PRCV (nasal samples). The RT-nested PCR assay was more sensitive than antigen-based as says on the basis of duration of virus detection in experimentally infected pigs and was directly applicable to nasal as well as fecal specimens from the field.