Xiao Wei, Biao Meng, Yan Li, Hong Peng, Xiangna Zhao
{"title":"Sensitive and rapid detection of Culex pipiens and Aedes albopictus","authors":"Xiao Wei, Biao Meng, Yan Li, Hong Peng, Xiangna Zhao","doi":"10.3389/finsc.2023.1015695","DOIUrl":null,"url":null,"abstract":"Background Culex pipiens and Aedes albopictus are closely related to human life, and transmit a variety of viruses, causing serious harm to human health. Cytochrome c oxidase I (COI) gene has been selected as a marker gene for studying phylogeny and molecular evolution of species and is also an effective molecular marker for studying the evolutionary mechanism and systematic reconstruction of diptera insects. Methods A loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of Cx. pipiens and Ae. albopictus were first described in this study. The experimental results were verified by real-time PCR. Results Our study showed the lower limit of sample concentration that can be detected by LAMP method is 0.5 pg/μl within 20 min for Cx. pipiens, and 1 pg/μl within 20 min for Ae. albopictus, which were more sensitive than PCR method. Validation tests with field samples showed LAMP method had good specificity and sensitivity and could identify the target species quickly and accurately. Conclusion The LAMP method developed in this study allowed the rapid and sensitive detection of Cx. pipiens and Ae. albopictus, which will be expected to be used for mass screening in batches of the field.","PeriodicalId":106657,"journal":{"name":"Frontiers in Insect Science","volume":"59 1","pages":"0"},"PeriodicalIF":0.0000,"publicationDate":"2023-03-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Frontiers in Insect Science","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.3389/finsc.2023.1015695","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
Background Culex pipiens and Aedes albopictus are closely related to human life, and transmit a variety of viruses, causing serious harm to human health. Cytochrome c oxidase I (COI) gene has been selected as a marker gene for studying phylogeny and molecular evolution of species and is also an effective molecular marker for studying the evolutionary mechanism and systematic reconstruction of diptera insects. Methods A loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of Cx. pipiens and Ae. albopictus were first described in this study. The experimental results were verified by real-time PCR. Results Our study showed the lower limit of sample concentration that can be detected by LAMP method is 0.5 pg/μl within 20 min for Cx. pipiens, and 1 pg/μl within 20 min for Ae. albopictus, which were more sensitive than PCR method. Validation tests with field samples showed LAMP method had good specificity and sensitivity and could identify the target species quickly and accurately. Conclusion The LAMP method developed in this study allowed the rapid and sensitive detection of Cx. pipiens and Ae. albopictus, which will be expected to be used for mass screening in batches of the field.
背景:库蚊和白纹伊蚊与人类生活密切相关,并传播多种病毒,对人类健康造成严重危害。细胞色素c氧化酶I (Cytochrome c oxidase I, COI)基因已被选为研究物种系统发育和分子进化的标记基因,也是研究双翅目昆虫进化机制和系统重建的有效分子标记。方法采用环介导等温扩增法(LAMP)快速、灵敏地检测Cx。库蚊和伊蚊。白纹伊蚊在本研究中首次被描述。通过实时荧光定量PCR对实验结果进行验证。结果Cx在20 min内的LAMP检测下限为0.5 pg/μl。20 min内1 pg/μl;白纹伊蚊,比PCR法更敏感。现场样品验证试验表明,LAMP方法具有良好的特异性和敏感性,能快速准确地鉴定出目标物种。结论本研究建立的LAMP方法能够快速、灵敏地检测Cx。库蚊和伊蚊。白纹伊蚊,预计将用于现场批量大规模筛查。