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Reverse transcriptase loop-mediated isothermal amplification for the detection of feline coronavirus

Author:
Stranieri, Angelica, Lauzi, Stefania, Giordano, Alessia, Paltrinieri, Saverio
Source:
Journal of virological methods 2017 v.243 pp. 105-108
ISSN:
0166-0934
Subject:
Felidae, Feline coronavirus, RNA, RNA-directed DNA polymerase, agarose, blood, cats, etiological agents, feces, feline infectious peritonitis, gels, lymph nodes, reverse transcription, reverse transcription loop-mediated isothermal amplification, viruses
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.
Agid:
5618098