Georgia Mataira, Fleur Francis, Jaimie Frazer, Robert Norton
{"title":"A comparison of DNA extraction methods used in the direct molecular detection of Burkholderia pseudomallei from blood","authors":"Georgia Mataira, Fleur Francis, Jaimie Frazer, Robert Norton","doi":"10.1093/trstmh/trae020","DOIUrl":null,"url":null,"abstract":"Background Melioidosis is caused by Burkholderia pseudomallei. Direct molecular detection from unamplified blood remains insensitive. Methods Three different extraction methods—QIAamp UCP Pathogen Mini Kit, High Pure PCR template and MagNA Pure Pathogen Universal—were trialled using spiked human ethylenediaminetetraacetic acid blood. A type III secretion system 1 (TTSS-1) polymerase chain reaction was used for detection. Results The QIAamp UCP Pathogen Mini Kit performed best, with a limit of detection of 1.5×102 cfu/ml. Conclusions It is planned to use the QIAamp UCP Pathogen Mini Kit to do a larger study on blood collected from patients with melioidosis.","PeriodicalId":501685,"journal":{"name":"Transactions of the Royal Society of Tropical Medicine & Hygiene","volume":"65 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2024-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Transactions of the Royal Society of Tropical Medicine & Hygiene","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1093/trstmh/trae020","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 Melioidosis is caused by Burkholderia pseudomallei. Direct molecular detection from unamplified blood remains insensitive. Methods Three different extraction methods—QIAamp UCP Pathogen Mini Kit, High Pure PCR template and MagNA Pure Pathogen Universal—were trialled using spiked human ethylenediaminetetraacetic acid blood. A type III secretion system 1 (TTSS-1) polymerase chain reaction was used for detection. Results The QIAamp UCP Pathogen Mini Kit performed best, with a limit of detection of 1.5×102 cfu/ml. Conclusions It is planned to use the QIAamp UCP Pathogen Mini Kit to do a larger study on blood collected from patients with melioidosis.