{"title":"用于病原菌单步富集的固体相的开发与表征","authors":"M. Archer, D. Stenger, B. Lin","doi":"10.2174/1874065000802010047","DOIUrl":null,"url":null,"abstract":"The identification of low abundance target nucleic acids in a complex matrix can be challenging due to the abundance background material. Current methods use two-step processes which are time consuming, prone to contamina- tion and usually limited to one pathogen. In this study we describe a single-step target-capture approach using magnetic microbeads with capture probes covalently attached through a phosphorus dendrimer linker. This approach was also used successfully for simultaneous capturing of two low abundance pathogenic nucleic acids present in a complex matrix (800- fold excess of background nucleic acids) by using a multi-pathogen solid phase. The thermal stability of the solid phase allows denaturation and capture to proceed sequentially and the recovery of the targets to be performed by heat denatura- tion without the risk of probe shedding. The critical variables involved in the development of the solid phase and the steps required for further optimization are discussed.","PeriodicalId":90363,"journal":{"name":"The open analytical chemistry journal","volume":"91 1","pages":"47-54"},"PeriodicalIF":0.0000,"publicationDate":"2008-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"3","resultStr":"{\"title\":\"Development and Characterization of a Solid Phase for Single-Step Enrichment of Pathogenic Targets\",\"authors\":\"M. Archer, D. Stenger, B. Lin\",\"doi\":\"10.2174/1874065000802010047\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"The identification of low abundance target nucleic acids in a complex matrix can be challenging due to the abundance background material. Current methods use two-step processes which are time consuming, prone to contamina- tion and usually limited to one pathogen. In this study we describe a single-step target-capture approach using magnetic microbeads with capture probes covalently attached through a phosphorus dendrimer linker. This approach was also used successfully for simultaneous capturing of two low abundance pathogenic nucleic acids present in a complex matrix (800- fold excess of background nucleic acids) by using a multi-pathogen solid phase. The thermal stability of the solid phase allows denaturation and capture to proceed sequentially and the recovery of the targets to be performed by heat denatura- tion without the risk of probe shedding. The critical variables involved in the development of the solid phase and the steps required for further optimization are discussed.\",\"PeriodicalId\":90363,\"journal\":{\"name\":\"The open analytical chemistry journal\",\"volume\":\"91 1\",\"pages\":\"47-54\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2008-05-09\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"3\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"The open analytical chemistry journal\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.2174/1874065000802010047\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"The open analytical chemistry journal","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.2174/1874065000802010047","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Development and Characterization of a Solid Phase for Single-Step Enrichment of Pathogenic Targets
The identification of low abundance target nucleic acids in a complex matrix can be challenging due to the abundance background material. Current methods use two-step processes which are time consuming, prone to contamina- tion and usually limited to one pathogen. In this study we describe a single-step target-capture approach using magnetic microbeads with capture probes covalently attached through a phosphorus dendrimer linker. This approach was also used successfully for simultaneous capturing of two low abundance pathogenic nucleic acids present in a complex matrix (800- fold excess of background nucleic acids) by using a multi-pathogen solid phase. The thermal stability of the solid phase allows denaturation and capture to proceed sequentially and the recovery of the targets to be performed by heat denatura- tion without the risk of probe shedding. The critical variables involved in the development of the solid phase and the steps required for further optimization are discussed.