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

IF 1.4 4区医学 Q3 PARASITOLOGY Korean Journal of Parasitology Pub Date : 2021-04-01 Epub Date: 2021-04-22 DOI:10.3347/kjp.2021.59.2.167

Yao-Dong Wu, Qi-Qi Wang, Meng Wang, Hany M Elsheikha, Xin Yang, Min Hu, Xing-Quan Zhu, Min-Jun Xu

{"title":"Development of a Lateral Flow Strip-Based Recombinase Polymerase Amplification Assay for the Detection of Haemonchus contortus in Goat Feces.","authors":"Yao-Dong Wu, Qi-Qi Wang, Meng Wang, Hany M Elsheikha, Xin Yang, Min Hu, Xing-Quan Zhu, Min-Jun Xu","doi":"10.3347/kjp.2021.59.2.167","DOIUrl":null,"url":null,"abstract":"<p><p>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.</p>","PeriodicalId":49938,"journal":{"name":"Korean Journal of Parasitology","volume":"59 2","pages":"167-171"},"PeriodicalIF":1.4000,"publicationDate":"2021-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/0c/b8/kjp-59-2-167.PMC8106983.pdf","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Korean Journal of Parasitology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3347/kjp.2021.59.2.167","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2021/4/22 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"PARASITOLOGY","Score":null,"Total":0}

引用次数: 1

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.

Abstract Image

查看原文

微信好友朋友圈 QQ好友复制链接

本刊更多论文

山羊粪便中扭曲血蜱横向流动条带重组酶扩增方法的建立。

在小型反刍动物中，血病仍然是一个重要的问题。本研究建立了重组酶聚合酶扩增(RPA)联合横向流动条带(LFS-RPA)快速检测山羊粪便中扭曲血蜱的方法。该试验使用引物和探针靶向ITS-2基因的特定序列。我们比较了LFS-RPA检测与PCR检测的性能。LFS-RPA的检出限为10 fg DNA，比PCR的最低检出限低10倍。在24份山羊粪便样本中，LFS-RPA法检测扭曲血蜱DNA的灵敏度为95.8%，而PCR的灵敏度为79.1%。LFS-RPA试验没有检测到其他相关蠕虫物种的DNA，并显示对山羊粪便中存在的抑制剂有足够的耐受性。综上所述，LFS-RPA法快速检测弓形虫具有较高的诊断准确性，值得进一步评价。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文去求助

来源期刊

Korean Journal of Parasitology PARASITOLOGY-

CiteScore

2.80

自引率

0.00%

发文量

审稿时长

1 months

期刊介绍： The Korean Journal of Parasitology is the official journal paperless, on-line publication after Vol. 53, 2015 of The Korean Society for Parasitology and Tropical Medicine. Abbreviated title is ‘Korean J Parasitol’. It was launched in 1963. It contains original articles, case reports, brief communications, reviews or mini-reviews, book reviews, and letters to the editor on parasites of humans and animals, vectors, host-parasite relationships, zoonoses, and tropical medicine. It is published bimonthly in February, April, June, August, October, and December each year. Supplement numbers are at times published. All of the manuscripts are peer-reviewed.