Nucleic acid-based differential diagnostic assays for feline coronavirus
| Autor: | Kyoung-Jin Yoon, Jae-Ik Han, Shien Young Kang, Ki Jeong Na |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
Veterinary Medicine
Feline coronavirus RT-PCR Cross Reactions Biology Cat Diseases medicine.disease_cause Sensitivity and Specificity Article Conserved sequence Coronavirus Canine Virology Canine coronavirus medicine Animals Multiplex Coronavirus Feline DNA Primers Coronavirus Reverse Transcriptase Polymerase Chain Reaction Transmissible gastroenteritis virus RNA virus biology.organism_classification Inosine Molecular Diagnostic Techniques Differentiation Cats Nucleic acid RNA Viral Primer (molecular biology) Coronavirus Infections Multiplex Polymerase Chain Reaction |
| Zdroj: | Journal of Virological Methods |
| ISSN: | 0166-0934 |
| DOI: | 10.1016/j.jviromet.2014.07.013 |
| Popis: | Highlights • A multiplex RT-PCR assay was developed to detect feline coronavirus. • To improve primer binding, inosine was used for primer design. • The assay could detect and differentiate FCoV, CCoV, and TGEV precisely. 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. |
| Databáze: | OpenAIRE |
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