Twist-Sensor: Novel microarrays for multiplex detection of drug resistance

DESCRIPTION provided by applicant Drug resistance is rapidly spreading among Gram negative bacteria seriously complicating the treatment of infected patients and increasing the risk of lethal outbreaks In particular carbapenem resistant Enterobacteriaceae CRE organisms associated with high mortality rate andgt have become prevalent worldwide During of long term acute care hospitals in the U S had at least one case of CRE Accurate and rapid identification of drug resistance is fundamental for proper treatment and prevention of nosocomial spread However microbiology laboratories have deficient tools for resistance diagnosis Phenotypic assays are the most common diagnostic methods but they are slow hours frequently inconclusive and may fail to detect carbapenemases Alternative methods based on nucleic acid detection exist however these methods are either time consuming and require trained personnel or only cover a limited number of resistance determinants Here we will develop a novel class of microarrays based on Twist Sensor technology for rapid accurate and multiplex detection of drug resistance Twist Sensor is a novel technology for nucleic acid detection based on single molecule DNA hybridization and supercoiling In this technique the hybridized targets are subjected to disrupting torsional stres which allows for rapid detection of multiple target sequences with extremely low background noise using a simple device We will demonstrate the capabilities of Twist Sensor technology by developing a microarray able to detect multiple carbapenemase genes in a procedure with minimum hands on time and with a total processing time of less than hours Aim will demonstrate that Twist Sensor technology is capable of detecting a resistance determinant in bacterial cultures without DNA amplification Aim will focus on developing a Twist Sensor detection mechanism that is easy to automate inexpensive and has high throughput We will use this novel microarray to develop a multiplex assay for detection of the most prevalent carbapenemases KPC NDM OXA VIM and IMP In order to optimize and characterize the functionality of the new microarray we will analyze bacterial culture samples of resistant isolates PUBLIC HEALTH RELEVANCE Drug resistance is rapidly spreading among Gram negative bacteria However diagnostic assays used in microbiology laboratories are slow and frequently inconclusive making treatment of infected patients difficult and increasing the risk of lethal outbreaks Here we will develop a novel class of DNA microarrays based on Twist Sensor technology for rapid accurate and multiplex detection of drug resistance