Development of a Lateral Flow Strip-Based Recombinase Polymerase Ampli cation Assay for the Detection of Haemonchus contortus in Goat Feces
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Haemonchosis remains a significant problem in small ruminants. In this study, the assay of recombinase poly- merase amplification (RPA) combined with the lateral flow strip (LFS-RPA) was established for the rapid detection of Hae- monchus 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 detec- tion of H. contortus and merits further evaluation.
Abstract
Haemonchosis remains a significant problem in small ruminants. In this study, the assay of recombinase poly- merase amplification (RPA) combined with the lateral flow strip (LFS-RPA) was established for the rapid detection of Hae- monchus 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 detec- tion of H. contortus and merits further evaluation.
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