Validation of the Right Now Test for Detection of spp. from Selected Environmental Surfaces Without Enrichment.

Right Now is a novel, enrichment-free molecular method for detection of spp. in swab samples from environmental surfaces. The test provides results in real time, indicating the current or recent presence of spp. in the environment. After sampling, the entire contents of the swab are subject to sample processing, releasing large quantities of target ribosomal RNA molecules into the lysate. A portion of the lysate is then tested using the ANSR for isothermal nucleic acid amplification assay. A study was conducted to validate the method for detection of spp. in swab samples from stainless steel and sealed concrete surfaces. In inclusivity testing, 60 of 60 spp. strains tested positive. In exclusivity testing, 31 of 31 nontarget bacterial strains tested negative. In LOD testing, the test was able to detect as few as 2 CFU of applied to a stainless steel surface. In matrix testing of inoculated stainless steel and sealed concrete surfaces, there were no statistically significant differences in method performance comparing the Right Now and U.S. Food and Drug Administration reference culture procedures as determined by probability of detection analysis. In robustness testing, modest changes to three assay operating parameters simultaneously did not significantly affect performance of the test. Results can be obtained in less than 1 h using the Right Now test, allowing food industry personnel to take immediate corrective action in the case of contamination incidents.