Recombinase polymerase amplification combined with lateral flow dipstick assay for rapid visual detection of A.simplex (s. s.) and A.pegreffii in sea foods
Anisakiasis is a food-borne parasitic disease mainly caused by the third stage of Anisakis simplex (s. s.) and Anisakis pegreffii. Traditional methods for detecting of Anisakis involve morphology identification such as visual inspection, enzyme digestion, and molecular methods based on PCR, but they...
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| Format: | Article |
| Language: | English |
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Elsevier
2024-04-01
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| Series: | Heliyon |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844024049740 |
| _version_ | 1827288757625159680 |
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| author | Xiaoming Wang Ting Xu Siling Ding Ye Xu Xingsheng Jin Feng Guan |
| author_facet | Xiaoming Wang Ting Xu Siling Ding Ye Xu Xingsheng Jin Feng Guan |
| author_sort | Xiaoming Wang |
| collection | DOAJ |
| description | Anisakiasis is a food-borne parasitic disease mainly caused by the third stage of Anisakis simplex (s. s.) and Anisakis pegreffii. Traditional methods for detecting of Anisakis involve morphology identification such as visual inspection, enzyme digestion, and molecular methods based on PCR, but they have certain limitations. In this study, the internal transcribed spacer 1 (ITS 1) regions of Anisakis were targeted to develop a visual screening method for detecting A. simplex (s. s.) and A. pegreffii in fish meat based on recombinase polymerase amplification (RPA) combined with lateral flow dipstick (LFD). Specific primers and probes were designed and optimized for temperature, reaction time, and detection threshold. LFD produced clear visual results that were easily identifiable after a consistent incubation of 10–20 min at 37 °C. The whole process of DNA amplification by RPA and readout by LFD did not exceed 30 min. In addition, the detection limit is up to 9.5 × 10−4 ng/μL, and the detection of the artificially contaminated samples showed that the developed assay can effectively and specifically detect A. simplex (s. s.) and A. pegreffii, which fully meet the market's requirements for fish food safety supervision. |
| first_indexed | 2024-04-24T11:38:08Z |
| format | Article |
| id | doaj.art-39c0d6d5708744ed9741be564f13eed6 |
| institution | Directory Open Access Journal |
| issn | 2405-8440 |
| language | English |
| last_indexed | 2024-04-24T11:38:08Z |
| publishDate | 2024-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Heliyon |
| spelling | doaj.art-39c0d6d5708744ed9741be564f13eed62024-04-10T04:29:11ZengElsevierHeliyon2405-84402024-04-01107e28943Recombinase polymerase amplification combined with lateral flow dipstick assay for rapid visual detection of A.simplex (s. s.) and A.pegreffii in sea foodsXiaoming Wang0Ting Xu1Siling Ding2Ye Xu3Xingsheng Jin4Feng Guan5College of Life Sciences, China Jiliang University, Hangzhou 310018, China; Zhejiang Museum of Natural History, Hangzhou 310018, ChinaCollege of Life Sciences, China Jiliang University, Hangzhou 310018, ChinaCollege of Life Sciences, China Jiliang University, Hangzhou 310018, ChinaCollege of Life Sciences, China Jiliang University, Hangzhou 310018, ChinaZhejiang Museum of Natural History, Hangzhou 310018, ChinaCollege of Life Sciences, China Jiliang University, Hangzhou 310018, China; Corresponding author.Anisakiasis is a food-borne parasitic disease mainly caused by the third stage of Anisakis simplex (s. s.) and Anisakis pegreffii. Traditional methods for detecting of Anisakis involve morphology identification such as visual inspection, enzyme digestion, and molecular methods based on PCR, but they have certain limitations. In this study, the internal transcribed spacer 1 (ITS 1) regions of Anisakis were targeted to develop a visual screening method for detecting A. simplex (s. s.) and A. pegreffii in fish meat based on recombinase polymerase amplification (RPA) combined with lateral flow dipstick (LFD). Specific primers and probes were designed and optimized for temperature, reaction time, and detection threshold. LFD produced clear visual results that were easily identifiable after a consistent incubation of 10–20 min at 37 °C. The whole process of DNA amplification by RPA and readout by LFD did not exceed 30 min. In addition, the detection limit is up to 9.5 × 10−4 ng/μL, and the detection of the artificially contaminated samples showed that the developed assay can effectively and specifically detect A. simplex (s. s.) and A. pegreffii, which fully meet the market's requirements for fish food safety supervision.http://www.sciencedirect.com/science/article/pii/S2405844024049740A. simplex (s. s.) and A. pegreffiiRecombinase polymerase amplificationLateral flow dipstickVisual detection |
| spellingShingle | Xiaoming Wang Ting Xu Siling Ding Ye Xu Xingsheng Jin Feng Guan Recombinase polymerase amplification combined with lateral flow dipstick assay for rapid visual detection of A.simplex (s. s.) and A.pegreffii in sea foods Heliyon A. simplex (s. s.) and A. pegreffii Recombinase polymerase amplification Lateral flow dipstick Visual detection |
| title | Recombinase polymerase amplification combined with lateral flow dipstick assay for rapid visual detection of A.simplex (s. s.) and A.pegreffii in sea foods |
| title_full | Recombinase polymerase amplification combined with lateral flow dipstick assay for rapid visual detection of A.simplex (s. s.) and A.pegreffii in sea foods |
| title_fullStr | Recombinase polymerase amplification combined with lateral flow dipstick assay for rapid visual detection of A.simplex (s. s.) and A.pegreffii in sea foods |
| title_full_unstemmed | Recombinase polymerase amplification combined with lateral flow dipstick assay for rapid visual detection of A.simplex (s. s.) and A.pegreffii in sea foods |
| title_short | Recombinase polymerase amplification combined with lateral flow dipstick assay for rapid visual detection of A.simplex (s. s.) and A.pegreffii in sea foods |
| title_sort | recombinase polymerase amplification combined with lateral flow dipstick assay for rapid visual detection of a simplex s s and a pegreffii in sea foods |
| topic | A. simplex (s. s.) and A. pegreffii Recombinase polymerase amplification Lateral flow dipstick Visual detection |
| url | http://www.sciencedirect.com/science/article/pii/S2405844024049740 |
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