Development of a highly sensitive point‐of‐care test for African swine fever that combines EZ‐Fast DNA extraction with LAMP detection: Evaluation using naturally infected swine whole blood samples from Vietnam

• Published in: Veterinary medicine and science Vol. 9; no. 3; pp. 1226 - 1233
• Main Authors: Ngan, Mai Thi, Thi My Le, Huynh, Xuan Dang, Vu, Thi Bich Ngoc, Trinh, Phan, Le Van, Thi Hoa, Nguyen, Quang Lam, Truong, Thi Lan, Nguyen, Notsu, Kosuke, Sekiguchi, Satoshi, Yamazaki, Yasuko, Yamazaki, Wataru
• Format: Journal Article
• Language: English
• Published: England John Wiley & Sons, Inc 01-05-2023; John Wiley and Sons Inc; Wiley
• Subjects: African swine fever; African Swine Fever - diagnosis; African Swine Fever Virus - genetics; Animals; Deoxyribonucleic acid; Disease; DNA; DNA, Viral; Epidemics; EZ‐Fast; field setting; Food security; Gene amplification; Genetic testing; Genomes; Hogs; Laboratories; Livestock; Livestock industry; loop‐mediated isothermal amplification; Mortality; Original; PIGS; Point-of-Care Testing; point‐of‐care test; Proteins; Quarantine; Sensitivity analysis; Swine; Swine Diseases; Vietnam; Viruses; Vietnam; Georgia; United States > US; Japan; Africa; EZ-Fast; African swine fever; loop-mediated isothermal amplification; field setting; point-of-care test
• ISSN: 2053-1095; 2053-1095
• DOI: 10.1002/vms3.1124

Background
While early detection and early containment are key to controlling the African swine fever (ASF) pandemic, the lack of practical testing methods for use in the field are a major barrier to achieving this feat.
Objectives
To describe the development of a rapid and sensitive point‐of‐care test (POCT) for ASF, and its evaluation using swine whole blood samples for field settings.
Methods
In total, 89 swine whole blood samples were collected from Vietnamese swine farms and were performed the POCT using a combination of crude DNA extraction and LAMP (loop‐mediated isothermal amplification) amplification.
Results
The POCT enabled crude DNA to be extracted from swine whole blood samples within 10 min at extremely low cost and with relative ease. The entire POCT required a maximum of 50 min from the beginning of DNA extraction to final judgment. Compared to a conventional real‐time PCR detection, the POCT showed a 1 log reduction in detection sensitivity, but comparable diagnostic sensitivity of 100% (56/56) and diagnostic specificity of 100% (33/33). The POCT was quicker and easier to perform and did not require special equipment.
Conclusions
This POCT is expected to facilitate early diagnosis and containment of ASF invasion into both regions in which it is endemic and eradicated.
The developed POCT is quicker and easier to perform and does not require special equipment. We validated a sensitive POCT for ASF in swine whole blood samples for field settings.This POCT is expected to facilitate early diagnosis and containment of ASF invasion into both regions in which it is endemic and eradicated.