The goal of this project over Phase I is to create and test the analytical validity of novel Invader mutation detection assays designed for applications specific to surgery and anesthesia. Invader technology, based on a highly specific enzyme-substrate reaction, enables direct analysis of genomic DNA without prior amplification, conferring advantages in rapidity, cost, and accuracy. The initial series of experiments will test 200 patients for 23 alleles of established clinical validity and utility at 12 loci predicting differences in patient responses to neuromuscular blocking agents (BChE, RYR1, CACNA1S), opioids and antiarrythmics (CYP2D6), nitrous oxide (MTHFR, MTR, MTRR, CBS), volatile anesthetics (RYR1, CACNA1s), and the risk and severity of sepsis (TNF-alpha, TNF-beta), and thrombosis (FVL, Prothrombin). The surgical- and anesthesia-specific Invader assays will be compared to conventional PCR-based methods with disparities resolved by DNA sequencing. Subsequent Phase II investigations will- add alleles of proven predictive value to multiplexed Invader patient panels designed for specific applications, and test whether these can favorably alter clinical outcomes. These data represent the necessary first steps in the assembly of high-throughput, automated perioperative genomic profiles incorporating hundreds of genotypes that can directly modulate risk and enhance the safety of all patients undergoing surgery. PROPOSED COMMERCIAL APPLICATIONS: Over Phases I and II, we aim to develop and validate a panel of Invader assays to detect clinically relevant mutations in the perioperative interval. Showing that this method can meet the precision, cost, and rapidity requirements needed for routine clinical testing, and improves patient safety, should make perioperative genomic profiling become routine in anesthetic and surgical care
