{"title":"基于RPA-LFD和RPA-CRISPR/Cas12a的pini疫霉快速检测技术的建立","authors":"Tingting Dai, Yufang Guo, Tongyue Wen, Sinong Yu, Yuan Tao, Zhuo Liu","doi":"10.3390/microorganisms13040863","DOIUrl":null,"url":null,"abstract":"<p><p><i>Phytophthora pini</i>, a globally dispersed plant pathogen, poses a significant threat to natural ecosystems and cultivated horticultural crops. Early and precise detection of <i>P. pini</i> is essential for effective disease management. This study focused on developing specific, rapid, and sensitive molecular diagnostic techniques to identify the pathogenic oomycete <i>P. pini</i>. We employed recombinase polymerase amplification with lateral flow device (RPA-LFD) and RPA combined with CRISPR/Cas12a. The RPA-LFD method can identify <i>P. pini</i> at concentrations as low as 10 pg/μL in 30 min, while the RPA-CRISPR/Cas12a approach can detect the pathogen at 1 pg/μL in approximately 50 min. These methods are highly effective in identifying disease caused by <i>P. pini</i> and provide a basis for future field detection, which may reduce the economic losses associated with this devastating disease.</p>","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":"13 4","pages":""},"PeriodicalIF":4.1000,"publicationDate":"2025-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12029582/pdf/","citationCount":"0","resultStr":"{\"title\":\"Establishment of a Rapid Detection Technique Based on RPA-LFD and RPA-CRISPR/Cas12a on <i>Phytophthora pini</i>.\",\"authors\":\"Tingting Dai, Yufang Guo, Tongyue Wen, Sinong Yu, Yuan Tao, Zhuo Liu\",\"doi\":\"10.3390/microorganisms13040863\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><i>Phytophthora pini</i>, a globally dispersed plant pathogen, poses a significant threat to natural ecosystems and cultivated horticultural crops. Early and precise detection of <i>P. pini</i> is essential for effective disease management. This study focused on developing specific, rapid, and sensitive molecular diagnostic techniques to identify the pathogenic oomycete <i>P. pini</i>. We employed recombinase polymerase amplification with lateral flow device (RPA-LFD) and RPA combined with CRISPR/Cas12a. The RPA-LFD method can identify <i>P. pini</i> at concentrations as low as 10 pg/μL in 30 min, while the RPA-CRISPR/Cas12a approach can detect the pathogen at 1 pg/μL in approximately 50 min. These methods are highly effective in identifying disease caused by <i>P. pini</i> and provide a basis for future field detection, which may reduce the economic losses associated with this devastating disease.</p>\",\"PeriodicalId\":18667,\"journal\":{\"name\":\"Microorganisms\",\"volume\":\"13 4\",\"pages\":\"\"},\"PeriodicalIF\":4.1000,\"publicationDate\":\"2025-04-10\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12029582/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Microorganisms\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.3390/microorganisms13040863\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Microorganisms","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.3390/microorganisms13040863","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
Establishment of a Rapid Detection Technique Based on RPA-LFD and RPA-CRISPR/Cas12a on Phytophthora pini.
Phytophthora pini, a globally dispersed plant pathogen, poses a significant threat to natural ecosystems and cultivated horticultural crops. Early and precise detection of P. pini is essential for effective disease management. This study focused on developing specific, rapid, and sensitive molecular diagnostic techniques to identify the pathogenic oomycete P. pini. We employed recombinase polymerase amplification with lateral flow device (RPA-LFD) and RPA combined with CRISPR/Cas12a. The RPA-LFD method can identify P. pini at concentrations as low as 10 pg/μL in 30 min, while the RPA-CRISPR/Cas12a approach can detect the pathogen at 1 pg/μL in approximately 50 min. These methods are highly effective in identifying disease caused by P. pini and provide a basis for future field detection, which may reduce the economic losses associated with this devastating disease.
期刊介绍：
Microorganisms (ISSN 2076-2607) is an international, peer-reviewed open access journal which provides an advanced forum for studies related to prokaryotic and eukaryotic microorganisms, viruses and prions. It publishes reviews, research papers and communications. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced. Electronic files and software regarding the full details of the calculation or experimental procedure, if unable to be published in a normal way, can be deposited as supplementary electronic material.
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