Academic Journal
Development of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Detecting Feline Coronavirus
| Title: | Development of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Detecting Feline Coronavirus |
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| Authors: | Witsanu Rapichai, Wichayet Saejung, Kotchaporn Khumtong, Chaiwat Boonkaewwan, Supansa Tuanthap, Peter A. Lieberzeit, Kiattawee Choowongkomon, Jatuporn Rattanasrisomporn |
| Source: | Animals, Vol 12, Iss 16, p 2075 (2022) |
| Publisher Information: | MDPI AG, 2022. |
| Publication Year: | 2022 |
| Collection: | LCC:Veterinary medicine LCC:Zoology |
| Subject Terms: | feline infectious peritonitis, reverse transcription loop-mediated isothermal amplification, molecular diagnosis, Veterinary medicine, SF600-1100, Zoology, QL1-991 |
| More Details: | Feline infectious peritonitis (FIP) is a worldwide fatal disease caused by a mutant feline coronavirus (FCoV). Simple and efficient molecular detection methods are needed. Here, sensitive, specific, rapid, and reliable colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed to detect the ORF1a/1b gene of FCoV from cats with suspected FIP using neutral red as an indicator. Novel LAMP primers were specifically designed based on the gene of interest. The isothermal assay could visually detect FCoV at 58 °C for 50 min. The RT-LAMP assay was highly specific and had no cross-reactivity with other related feline viruses. The detection limit of FCoV detection by RT-LAMP was 20 fg/µL. A blind clinical test (n = 81) of the developed RT-LAMP procedure was in good agreement with the conventional PCR method. In the light of its performance specificity, sensitivity, and easy visualization, this neutral-red-based RT-LAMP approach would be a fruitful alternative molecular diagnostic tool for veterinary inspection of FCoV when combined with nucleotide sequencing or specific PCR to affirm the highly virulent FIP-associated FCoV. |
| Document Type: | article |
| File Description: | electronic resource |
| Language: | English |
| ISSN: | 2076-2615 |
| Relation: | https://www.mdpi.com/2076-2615/12/16/2075; https://doaj.org/toc/2076-2615 |
| DOI: | 10.3390/ani12162075 |
| Access URL: | https://doaj.org/article/748f26b852a64de7adb625af633e5c4d |
| Accession Number: | edsdoj.748f26b852a64de7adb625af633e5c4d |
| Database: | Directory of Open Access Journals |
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| ISSN: | 20762615 |
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| DOI: | 10.3390/ani12162075 |
| Published in: | Animals |
| Language: | English |