title: Reverse transcriptase loop-mediated isothermal amplification for the detection of feline coronavirus.
abstract: The Feline coronavirus (FCoV) is the etiological agent of feline infectious peritonitis (FIP), a lethal disease of felids. The role of molecular methods is controversial for the diagnosis of FIP, while essential for the identification of the shedders. Thus, a fast and inexpensive method for the detection of FCoV could be beneficial, especially in multicat environments. A reverse transcription loop mediated isothermal amplification (RT-LAMP) assay was developed. RNA extraction and RT-nPCR for FCoV were performed on thirty-two samples (11 faeces, 9 blood, 8 effusions, and 4 lymph nodes) collected from 27 cats. Six RT-LAMP primers were designed from the same conserved region of RT-nPCR, and the assay was run at 63°C for one hour. Results were evaluated through both agarose gel run and hydroxynapthol blue (HNB) dye and then compared with RT-nPCR results for the assessment of sensitivity and specificity. The overall specificity was 100%, but the sensitivity was 50% and 54.5% for agarose gel and HNB respectively. Therefore, RT-LAMP seems optimal to confirm the presence of the virus, but not applicable to exclude it.
authors.affiliation: Department of Veterinary Medicine, University of Milan, Milan, Italy; Veterinary Teaching Hospital, University of Milan, Milan, Italy. Electronic address: angelica.stranieri@unimi.it.
authors.forename: Angelica
authors.initials: A
authors.lastname: Stranieri
authors.affiliation: Department of Veterinary Medicine, University of Milan, Milan, Italy; Veterinary Teaching Hospital, University of Milan, Milan, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Lauzi
authors.affiliation: Department of Veterinary Medicine, University of Milan, Milan, Italy; Veterinary Teaching Hospital, University of Milan, Milan, Italy.
authors.forename: Alessia
authors.initials: A
authors.lastname: Giordano
authors.affiliation: Department of Veterinary Medicine, University of Milan, Milan, Italy; Veterinary Teaching Hospital, University of Milan, Milan, Italy.
authors.forename: Saverio
authors.initials: S
authors.lastname: Paltrinieri
mesh_terms: D000818:Animals; D002415:Cats; D016765:Coronavirus, Feline / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D016766:Feline Infectious Peritonitis / Q000175:diagnosis* / Q000821:virology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D013696:Temperature
chemical_list: D017931:DNA Primers; D012367:RNA, Viral
keywords: Feline infectious peritonitis; Hydroxynaphtol blue; Isothermal gene amplification