TY - JOUR
T1 - Innovative Colorimetric Neutral Red-Based Loop-Mediated Isothermal Amplification (NR-LAMP) Assay: Transforming Rapid and Affordable Feline Leukemia Virus Detection
AU - Rapichai, Witsanu
AU - Khamsingnok, Piyamat
AU - Wachirachaikarn, Anyalak
AU - Laodim, Thawee
AU - Dong, Hieu Van
AU - Meecharoen, Nianrawan
AU - Ratanabunyong, Siriluk
AU - Khaoiam, Thanawat
AU - Tuanthap, Supansa
AU - Rattanasrisomporn, Amonpun
AU - Pairor, Selapoom
AU - Choowongkomon, Kiattawee
AU - Tansakul, Natthasit
AU - Lieberzeit, Peter A.
AU - Rattanasrisomporn, Jatuporn
N1 - Publisher Copyright:
© 2025 by the authors.
Accession Number
WOS:001646783400001
PubMed ID
41465225
PY - 2025/12
Y1 - 2025/12
N2 - Feline leukemia virus (FeLV) is a retrovirus that globally affects both domestic and wild cats, leading to the development of leukemia, lymphoma, and immunosuppression. However, it is important to note that the daily antigen test may yield false negative results. In this study, we successfully developed the first colorimetric loop-mediated isothermal amplification (LAMP) associated with neutral red (NR-LAMP) for the detection of FeLV. The NR-LAMP assay exhibited high sensitivity and efficiency compared to the routine polymerase chain reaction (PCR) reference method. To ensure specificity, a novel LAMP primer set was custom-designed based on the pol gene of multiple FeLV strains, which resulted in no cross-amplification with other feline viruses. Under the optimized isothermal amplification conditions at 61 °C for 40 min, the NR-LAMP assay achieved a detection limit of 100 copies/µL. Using a blind clinical test involving 98 samples, the NR-LAMP assay demonstrated perfect agreement with the reference PCR method, providing a sensitivity of 97.3% and a specificity of 100%. This proposed NR-LAMP assay surpasses other related approaches in terms of sensitivity, efficiency, and cost-effectiveness. Consequently, the colorimetric NR-LAMP reaction serves as a robust and convenient diagnostic tool for the inspection of FeLV, offering an alternative molecular method for future clinical applications and commercial utilization.
AB - Feline leukemia virus (FeLV) is a retrovirus that globally affects both domestic and wild cats, leading to the development of leukemia, lymphoma, and immunosuppression. However, it is important to note that the daily antigen test may yield false negative results. In this study, we successfully developed the first colorimetric loop-mediated isothermal amplification (LAMP) associated with neutral red (NR-LAMP) for the detection of FeLV. The NR-LAMP assay exhibited high sensitivity and efficiency compared to the routine polymerase chain reaction (PCR) reference method. To ensure specificity, a novel LAMP primer set was custom-designed based on the pol gene of multiple FeLV strains, which resulted in no cross-amplification with other feline viruses. Under the optimized isothermal amplification conditions at 61 °C for 40 min, the NR-LAMP assay achieved a detection limit of 100 copies/µL. Using a blind clinical test involving 98 samples, the NR-LAMP assay demonstrated perfect agreement with the reference PCR method, providing a sensitivity of 97.3% and a specificity of 100%. This proposed NR-LAMP assay surpasses other related approaches in terms of sensitivity, efficiency, and cost-effectiveness. Consequently, the colorimetric NR-LAMP reaction serves as a robust and convenient diagnostic tool for the inspection of FeLV, offering an alternative molecular method for future clinical applications and commercial utilization.
KW - colorimetric LAMP
KW - cost-effective clinical application
KW - feline leukemia virus (FeLV)
KW - neutral red
KW - pol gene
UR - https://www.scopus.com/pages/publications/105025914071
U2 - 10.3390/ijms262411793
DO - 10.3390/ijms262411793
M3 - Article
AN - SCOPUS:105025914071
SN - 1661-6596
VL - 26
JO - International Journal of Molecular Sciences
JF - International Journal of Molecular Sciences
IS - 24
M1 - 11793
ER -