Detection of feline immunodeficiency virus by neutral red-based loop-mediated isothermal amplification assay

Vet World Vol.17 January-2024 Article-8

Research Article

Veterinary World, 17(1): 72-81

https://doi.org/10.14202/vetworld.2024.72-81

Wichayet Saejung^1,2, Kotchaporn Khumtong^1,2, Witsanu Rapichai^2,3, Siriluk Ratanabunyong³, Amonpun Rattanasrisomporn⁴, Kiattawee Choowongkomon³, Oumaporn Rungsuriyawiboon⁵, and Jatuporn Rattanasrisomporn^1,2

1. Graduate Program in Animal Health and Biomedical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand.
2. Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand.
3. Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok, Thailand.
4. Interdisciplinary of Genetic Engineering and Bioinformatics, Graduate School, Kasetsart University, Bangkok, Thailand.
5. Department of Veterinary Technology, Faculty of Veterinary Technology, Kasetsart University, Bangkok, Thailand.

Background and Aim: Feline immunodeficiency virus (FIV) is a retroviral pathogen globally responsible for immunodeficiency disease in cats. However, the current diagnosis based on antibody detection has limitations and can also produce false-positive results. This study aimed to develop a one-pot loop-mediated isothermal amplification (LAMP) process integrated with neutral red (NR-LAMP) assay for detection of FIV proviral DNA.

Materials and Methods: We developed a one-pot, gag gene-based NR-LAMP for convenient, rapid, specific, and sensitive colorimetric inspection of FIV proviral DNA.

Results: The developed NR-LAMP was capable of amplifying at an optimum temperature of 65°C for 40 min. No cross-amplification was detected between FIV and other feline viruses tested, indicating the high specificity (98.44%) of the novel FIV-LAMP primer. Our NR-LAMP assay has a detection limit of 4.2 × 101 copies/μL. A total of 80 clinical samples with a background of FIV infection were collected and tested using the proposed method. The NR-LAMP assay showed a high sensitivity of 100% compared to conventional polymerase chain reaction assay.

Conclusion: These results support the suitability of NR-LAMP as a potential future alternative clinical molecular approach for further use in the diagnosis of FIV-infected cats. Keywords: feline immunodeficiency virus, loop-mediated isothermal amplification, molecular diagnosis, neutral red.

Keywords: feline immunodeficiency virus, loop-mediated isothermal amplification, molecular diagnosis, neutral red.

How to cite this article: Saejung W, Khumtong K, Rapichai W, Ratanabunyong S, Rattanasrisomporn A, Choowongkomon K, Rungsuriyawiboon O, and Rattanasrisomporn J (2024) Detection of feline immunodeficiency virus by neutral red-based loop-mediated isothermal amplification assay, Veterinary World, 17(1): 72-81.

Received: 20-09-2023 Accepted: 13-12-2023 Published online: 08-01-2024

Corresponding author: Jatuporn Rattanasrisomporn E-mail: fvetjpn@ku.ac.th

DOI: 10.14202/vetworld.2024.72-81

Copyright: Saejung, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.