Open Access
Research (Published online: 08-01-2024)
8. Detection of feline immunodeficiency virus by neutral red-based loop-mediated isothermal amplification assay
Wichayet Saejung, Kotchaporn Khumtong, Witsanu Rapichai, Siriluk Ratanabunyong, Amonpun Rattanasrisomporn, Kiattawee Choowongkomon, Oumaporn Rungsuriyawiboon, and Jatuporn Rattanasrisomporn
Veterinary World, 17(1): 72-81

Wichayet Saejung: Graduate Program in Animal Health and Biomedical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand; Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand.
Kotchaporn Khumtong: Graduate Program in Animal Health and Biomedical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand; Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand.
Witsanu Rapichai: Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand; Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok, Thailand.
Siriluk Ratanabunyong: Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok, Thailand.
Amonpun Rattanasrisomporn: Interdisciplinary of Genetic Engineering and Bioinformatics, Graduate School, Kasetsart University, Bangkok, Thailand.
Kiattawee Choowongkomon: Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok, Thailand.
Oumaporn Rungsuriyawiboon: Department of Veterinary Technology, Faculty of Veterinary Technology, Kasetsart University, Bangkok, Thailand.
Jatuporn Rattanasrisomporn: Graduate Program in Animal Health and Biomedical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand; Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand.

doi: 10.14202/vetworld.2024.72-81

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

Corresponding author: Jatuporn Rattanasrisomporn

E-mail: fvetjpn@ku.ac.th

Citation: 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.
Abstract

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.