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Development of a Lateral Flow Strip-Based Recombinase Polymerase Amplification Assay for the Detection of Haemonchus contortus in Goat Feces

Korean Journal of Parasitology 2021년 59권 2호 p.167 ~ 171
Wu Yao-Dong, Wang Qi-Qi, Wang Meng, Elsheikha Hany M., Yang Xin, Hu Min, Zhu Xing-Quan, Xu Min-Jun,
소속 상세정보
 ( Wu Yao-Dong ) - South China Agricultural University College of Marine Sciences
 ( Wang Qi-Qi ) - Chinese Academy of Agricultural Sciences Lanzhou Veterinary Research Institute
 ( Wang Meng ) - Chinese Academy of Agricultural Sciences Lanzhou Veterinary Research Institute
 ( Elsheikha Hany M. ) - University of Nottingham School of Veterinary Medicine and Science
 ( Yang Xin ) - South China Agricultural University College of Veterinary Medicine
 ( Hu Min ) - Huazhong Agricultural University College of Veterinary Medicine
 ( Zhu Xing-Quan ) - Chinese Academy of Agricultural Sciences Lanzhou Veterinary Research Institute
 ( Xu Min-Jun ) - South China Agricultural University College of Marine Sciences

Abstract


Haemonchosis remains a significant problem in small ruminants. In this study, the assay of recombinase polymerase amplification (RPA) combined with the lateral flow strip (LFS-RPA) was established for the rapid detection of Haemonchus contortus in goat feces. The assay used primers and a probe targeting a specific sequence in the ITS-2 gene. We compared the performance of the LFS-RPA assay to a PCR assay. The LFS-RPA had a detection limit of 10 fg DNA, which was 10 times less compared to the lowest detection limit obtained by PCR. Out of 24 goat fecal samples, LFS-RPA assay detected H. contortus DNA with 95.8% sensitivity, compared to PCR, 79.1% sensitivity. LFS-RPA assay did not detect DNA from other related helminth species and demonstrated an adequate tolerance to inhibitors present in the goat feces. Taken together, our results suggest that LFS-RPA assay had a high diagnostic accuracy for the rapid detection of H. contortus and merits further evaluation.

키워드

Haemonchus contortus; goat; recombinase polymerase amplification; lateral flow strip; rapid detection

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