Wang Aiying, Luo Ju, Wang Cilin, Hou Yuxuan, Yang Baojun, Tang Jian, Liu Shuhua
{"title":"重组酶辅助扩增-侧向流双液相色谱法在水稻上的建立及应用","authors":"Wang Aiying, Luo Ju, Wang Cilin, Hou Yuxuan, Yang Baojun, Tang Jian, Liu Shuhua","doi":"10.1007/s13313-023-00918-8","DOIUrl":null,"url":null,"abstract":"<div><p><i>Pantoea ananatis</i> is a major pathogen that causes the new bacterial blight in rice, and its symptoms very similar to rice bacterial blight. Therefore, there is a dire need for an accurate and rapid method for detecting <i>P. ananatis</i>. In this study, an early and rapid visual detection method for <i>P. ananatis</i> was established. Using GyrB gene as the target sequence, an innovative recombinase-aided amplification detection system integrated with a lateral flow dipstick (RAA-LFD) was constructed. The optimized RAA-LFD detection method can be initiated at body temperature and does not rely on precise instruments. It does not require DNA extraction and can be used directly with plant tissue fluids. The results can be visualized after 10 minutes of amplification. The specificity and sensitivity tests showed that the RAA-LFD method could detect <i>P. ananatis,</i> whereas other common plant pathogens were not detected, and its detection sensitivity for <i>P. ananatis</i> DNA reached 100 copies/µL. The detection of diseased tissues indicated that this method could accurately detect <i>P. ananatis</i> in artificially inoculated rice tissues in the early stages of infection before symptoms. The RAA-LFD detection system established in this study is simple and fast, with visual results, excellent specificity, and high sensitivity. It is semi-quantitative and should be used for the early detection and rapid field diagnosis of new leaf blight, which provides technical support for the early warning and real-time detection of field samples.</p></div>","PeriodicalId":8598,"journal":{"name":"Australasian Plant Pathology","volume":null,"pages":null},"PeriodicalIF":0.9000,"publicationDate":"2023-05-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":"{\"title\":\"Establishment and application of the Recombinase-Aided Amplification-Lateral Flow Dipstick detection method for Pantoea ananatis on rice\",\"authors\":\"Wang Aiying, Luo Ju, Wang Cilin, Hou Yuxuan, Yang Baojun, Tang Jian, Liu Shuhua\",\"doi\":\"10.1007/s13313-023-00918-8\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p><i>Pantoea ananatis</i> is a major pathogen that causes the new bacterial blight in rice, and its symptoms very similar to rice bacterial blight. Therefore, there is a dire need for an accurate and rapid method for detecting <i>P. ananatis</i>. In this study, an early and rapid visual detection method for <i>P. ananatis</i> was established. Using GyrB gene as the target sequence, an innovative recombinase-aided amplification detection system integrated with a lateral flow dipstick (RAA-LFD) was constructed. The optimized RAA-LFD detection method can be initiated at body temperature and does not rely on precise instruments. It does not require DNA extraction and can be used directly with plant tissue fluids. The results can be visualized after 10 minutes of amplification. The specificity and sensitivity tests showed that the RAA-LFD method could detect <i>P. ananatis,</i> whereas other common plant pathogens were not detected, and its detection sensitivity for <i>P. ananatis</i> DNA reached 100 copies/µL. The detection of diseased tissues indicated that this method could accurately detect <i>P. ananatis</i> in artificially inoculated rice tissues in the early stages of infection before symptoms. The RAA-LFD detection system established in this study is simple and fast, with visual results, excellent specificity, and high sensitivity. It is semi-quantitative and should be used for the early detection and rapid field diagnosis of new leaf blight, which provides technical support for the early warning and real-time detection of field samples.</p></div>\",\"PeriodicalId\":8598,\"journal\":{\"name\":\"Australasian Plant Pathology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.9000,\"publicationDate\":\"2023-05-18\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Australasian Plant Pathology\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://link.springer.com/article/10.1007/s13313-023-00918-8\",\"RegionNum\":4,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"PLANT SCIENCES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Australasian Plant Pathology","FirstCategoryId":"97","ListUrlMain":"https://link.springer.com/article/10.1007/s13313-023-00918-8","RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"PLANT SCIENCES","Score":null,"Total":0}
Establishment and application of the Recombinase-Aided Amplification-Lateral Flow Dipstick detection method for Pantoea ananatis on rice
Pantoea ananatis is a major pathogen that causes the new bacterial blight in rice, and its symptoms very similar to rice bacterial blight. Therefore, there is a dire need for an accurate and rapid method for detecting P. ananatis. In this study, an early and rapid visual detection method for P. ananatis was established. Using GyrB gene as the target sequence, an innovative recombinase-aided amplification detection system integrated with a lateral flow dipstick (RAA-LFD) was constructed. The optimized RAA-LFD detection method can be initiated at body temperature and does not rely on precise instruments. It does not require DNA extraction and can be used directly with plant tissue fluids. The results can be visualized after 10 minutes of amplification. The specificity and sensitivity tests showed that the RAA-LFD method could detect P. ananatis, whereas other common plant pathogens were not detected, and its detection sensitivity for P. ananatis DNA reached 100 copies/µL. The detection of diseased tissues indicated that this method could accurately detect P. ananatis in artificially inoculated rice tissues in the early stages of infection before symptoms. The RAA-LFD detection system established in this study is simple and fast, with visual results, excellent specificity, and high sensitivity. It is semi-quantitative and should be used for the early detection and rapid field diagnosis of new leaf blight, which provides technical support for the early warning and real-time detection of field samples.
期刊介绍:
Australasian Plant Pathology presents new and significant research in all facets of the field of plant pathology. Dedicated to a worldwide readership, the journal focuses on research in the Australasian region, including Australia, New Zealand and Papua New Guinea, as well as the Indian, Pacific regions.
Australasian Plant Pathology is the official journal of the Australasian Plant Pathology Society.