Angione Stephanie L, Sarma Aartik A, Novikov Aleksey, Seward Leah, Fieber Jennifer H, Mermel Leonard A, Tripathi Anubhav
Center for Biomedical Engineering, School of Engineering, Brown University, Providence, Rhode Island.
Warren Alpert Medical School, Brown University, Providence, Rhode Island.
J Mol Diagn. 2014 Mar;16(2):244-52. doi: 10.1016/j.jmoldx.2013.11.006. Epub 2014 Jan 13.
This proof-of-concept study demonstrates the application of a novel nucleic acid detection platform to detect Clostridium difficile in subjects presenting with acute diarrheal symptoms. This method amplifies three genes associated with C. difficile infection, including genes and deletions (cdtB and tcdC) associated with hypervirulence attributed to the NAP1/027/BI strain. Amplification of DNA from the tcdB, tcdC, and cdtB genes was performed using a droplet-based sandwich platform with quantitative real-time PCR in microliter droplets to detect and identify the amplified fragments of DNA. The device and identification system are simple in design and can be integrated as a point-of-care test to help rapidly detect and identify C. difficile strains that pose significant health threats in hospitals and other health-care communities.
这项概念验证研究展示了一种新型核酸检测平台在检测出现急性腹泻症状的受试者中艰难梭菌的应用。该方法扩增了与艰难梭菌感染相关的三个基因，包括与NAP1/027/BI菌株所致高毒力相关的基因和缺失（cdtB和tcdC）。使用基于微滴的夹心平台和微升液滴中的定量实时PCR对tcdB、tcdC和cdtB基因的DNA进行扩增，以检测和鉴定扩增的DNA片段。该设备和识别系统设计简单，可整合为即时检测，以帮助快速检测和识别在医院及其他医疗社区中构成重大健康威胁的艰难梭菌菌株。
