title: Nucleic acid-based differential diagnostic assays for feline coronavirus.
abstract: Feline coronavirus (FCoV) is a pleomorphic, enveloped, positive-sense single-stranded RNA virus. Owing to the differences in its genotype, FCoV belongs to a separate clade along with other viruses, such as transmissible gastroenteritis virus (TGEV) and canine coronavirus (CCoV), which can be isolated from cats. In this study, a PCR assay was developed to differentiate these coronaviruses concurrently. Multiplex differential RT-PCR was performed with primers based on the highly conserved coronavirus membrane protein. Three primer sets were designed: a primer pair (S1 and S2) that can bind to conserved sequences in all target coronaviruses, a CCoV-specific primer (S3), and a TGEV-specific primer (S4). Because of the high sequence homology among FCoV, CCoV, and TGEV, a nucleotide preceding the last pair of dissimilar nucleotides in S3 and S4 was substituted with an inosine to allow primer binding. This assay could detect and differentiate FCoV (n=7), CCoV (n=4), and TGEV (n=8) precisely and did not show any cross-reactivity with other pathogens. These results suggest that this molecular approach provides a rapid and reliable way to detect FCoV, especially in feline clinical specimens.
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju 361-763, South Korea; Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA.
authors.forename: Jae-Ik
authors.initials: JI
authors.lastname: Han
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju 361-763, South Korea.
authors.forename: Shien-Young
authors.initials: SY
authors.lastname: Kang
authors.affiliation: Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA.
authors.forename: Kyoung-Jin
authors.initials: KJ
authors.lastname: Yoon
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju 361-763, South Korea. Electronic address: sigol@cbnu.ac.kr.
authors.forename: Ki-Jeong
authors.initials: KJ
authors.lastname: Na
mesh_terms: D000818:Animals; D002371:Cat Diseases / Q000175:diagnosis* / Q000821:virology; D002415:Cats; D018352:Coronavirus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D017939:Coronavirus, Canine / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D016765:Coronavirus, Feline / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D003429:Cross Reactions; D017931:DNA Primers / Q000235:genetics; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D005760:Transmissible gastroenteritis virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D014730:Veterinary Medicine / Q000379:methods*
chemical_list: D017931:DNA Primers; D012367:RNA, Viral
keywords: Canine coronavirus; Differentiation; Feline coronavirus; Inosine; RT-PCR; Transmissible gastroenteritis virus
references.citation: Vet Clin North Am Small Anim Pract. 2005 Jan;35(1):39-79, vi
references.pmid: 15627627
references.citation: Virology. 1992 Nov;191(1):134-40
references.pmid: 1329312
references.citation: Vet Microbiol. 2004 Mar 26;99(1):31-42
references.pmid: 15019109
references.citation: J Clin Microbiol. 1983 Feb;17(2):202-17
references.pmid: 6300184
references.citation: Vet Microbiol. 1998 Jul;62(3):193-205
references.pmid: 9791867
references.citation: Am J Vet Res. 1981 Mar;42(3):368-77
references.pmid: 6267960
references.citation: Vet Microbiol. 2000 Jul 31;75(2):127-33
references.pmid: 10889403
references.citation: J Clin Microbiol. 2002 Nov;40(11):4126-30
references.pmid: 12409385
references.citation: J Virol Methods. 2005 Mar;124(1-2):111-6
references.pmid: 15664058
references.citation: Am J Vet Res. 1979 Oct;40(10):1487-92
references.pmid: 230758
references.citation: J Virol Methods. 1999 Jun;80(1):11-5
references.pmid: 10403671
references.citation: J Gen Virol. 2003 Oct;84(Pt 10):2735-2744
references.pmid: 13679608
references.citation: J Vet Diagn Invest. 2000 Jul;12(4):385-8
references.pmid: 10907874
references.citation: Proc Natl Acad Sci U S A. 1998 Apr 14;95(8):4258-63
references.pmid: 9539724
references.citation: J Clin Microbiol. 2006 Mar;44(3):916-22
references.pmid: 16517876
references.citation: Nucleic Acids Res. 1988 Nov 25;16(22):10932
references.pmid: 2462716
references.citation: J Vet Diagn Invest. 2011 Sep;23(5):951-5
references.pmid: 21908353
references.citation: J Feline Med Surg. 2007 Oct;9(5):369-72
references.pmid: 17478116
references.citation: J Virol. 1989 Jan;63(1):436-40
references.pmid: 2521188
references.citation: J Am Vet Med Assoc. 2005 May 1;226(9):1503-7
references.pmid: 15882000
references.citation: Clin Microbiol Rev. 2000 Oct;13(4):559-70
references.pmid: 11023957
references.citation: Am J Vet Res. 1992 Sep;53(9):1500-6
references.pmid: 1329586
references.citation: J Virol Methods. 2009 Dec;162(1-2):64-8
references.pmid: 19646484
references.citation: Emerg Infect Dis. 2013 Jul;19(7):1066-73
references.pmid: 23763835
references.citation: Emerg Infect Dis. 2009 Sep;15(9):1445-52
references.pmid: 19788813