Molecular diagnostics in a teacup:Non-Instrumented Nucleic Acid Amplification(NINA) for rapid,low cost detection of Salmonella enterica

作     者：KUBOTA Ryo LABARRE Paul WEIGL Bernhard H LI Yong HAYDOCK Paul JENKINS Daniel M 

作者机构：Molecular Biosciences and BioengineeringUniversity of Hawaii at ManoaHonoluluHI 96822USA DiageneticsIncHonoluluHI 96822USA Program for Appropriate Technologies in Health(PATH)SeattleWA 98109USA Human NutritionFoodand Animal ScienceUniversity of Hawaii at ManoaHonoluluHI 96822USA Blood Cell Storage Inc(BCSI)SeattleWA 98109USA 

出 版 物：《Chinese Science Bulletin》 (Chinese Science Bulletin)

年 卷 期：2013年第58卷第10期

页      面：1162-1168页

核心收录：

学科分类：0710[理学-生物学] 1007[医学-药学(可授医学、理学学位)] 100705[医学-微生物与生化药学] 07[理学] 071005[理学-微生物学] 10[医学] 

基　　金：Financial support at UH was provided by USDA-Hatch project HA-517 NINA concepts and prototypes developed at PATH were supported by grants under the Health Innovations Portfolio,which is supported by the United States Agency for International Development,private foundations,and individual donors 

主　　题：沙门氏菌污染 检测设备 分子诊断 NINA 核酸扩增 成本低 反应温度 仪表 

摘      要：We report on the use of a novel non-instrumented platform to enable a Loop Mediated isothermal Amplification(LAMP) based assay for Salmonella *** energy is provided by addition of a small amount(150 g) of boiling water,and the reaction temperature is regulated by storing latent energy at the melting temperature of a lipid-based engineered phase change *** classification of the reaction is achieved without opening the reaction tube by observing the fluorescence of sequence-specific FRET-based assimilating probes with a simple handheld *** or above 22℃ ambient temperature the non-instrumented devices could maintain reactions above a threshold temperature of 61℃ for over 90 min-significantly longer than the 60 min reaction *** the simple format,detection limits were less than 20 genome copies for reactions run at ambient temperatures ranging from 8 to 36℃.When used with a pre-enrichment step and non-instrumented DNA extraction device,trace contaminations of Salmonella in milk close to 1 CFU/mL could be reliably *** findings illustrate that the non-instrumented amplification approach is a simple,viable,low-cost alternative for field-based food and agricultural diagnostics or clinical applications in developing countries.