Rapid detection of alveolar echinococcosis in hepatic nodules of horses by recombinase polymerase amplification assay

Tatsuro Hifumi; Tetsuya Tanaka; Miho Sato; Kohei Akioka; Chiaki Fujimata; Noriaki Miyoshi

doi:10.1016/j.vas.2023.100291

Rapid detection of alveolar echinococcosis in hepatic nodules of horses by recombinase polymerase amplification assay

Vet Anim Sci. 2023 Mar 2:20:100291. doi: 10.1016/j.vas.2023.100291. eCollection 2023 Jun.

Authors

Tatsuro Hifumi¹, Tetsuya Tanaka², Miho Sato¹, Kohei Akioka³, Chiaki Fujimata³, Noriaki Miyoshi¹

Affiliations

¹ Laboratory of Veterinary Histopathology, Joint Faculty of Veterinary Medicine, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.
² Laboratory of Infectious Diseases, Joint Faculty of Veterinary Medicine, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.
³ Kumamoto Prefectural Meat Inspection Center, 1314 Sosaki, Shichijo-machi, Kikuchi, Kumamoto 861-1344, Japan.

Abstract

Alveolar echinococcosis in slaughtered horses remains a public health issue. This study aimed to develop a Recombinase polymerase amplification (RPA) assay targeting the mitochondrial NADH dehydrogenase subunit 5 (Nad5) gene of Echinococcus multilocularis for the rapid detection of equine alveolar echinococcosis. Thirty-six hepatic solid nodules obtained from each horse (n = 36) were evaluated based on histopathological examination and Nad5-targeted PCR and then submitted to the RPA assay. The results of the developed RPA assay were 94.4% consistent with those of Nad5 PCR and Cohen's kappa coefficient value was 0.89 statistically, indicating high agreement. In addition, the RPA assay using the plasmid samples was one hundredfold more sensitive than PCR testing. Consequently, these results suggest that the performance of the RPA assay developed in this study is equal to that of conventional PCR testing.

Keywords: Alveolar echinococcosis; HE, hematoxylin and eosin; Horse; LAMP, loop-mediated isothermal amplification; Nad5, mitochondrial NADH dehydrogenase subunit 5; PAS, periodic acid-Schiff; PCR, polymerase chain reaction; RPA, recombinase polymerase amplification; Recombinase polymerase amplification.