Potential testing of reprocessing procedures by real-time polymerase chain reaction: A multicenter study of colonoscopy devices

Publication date: Available online 25 September 2017
Source:American Journal of Infection Control
Author(s): Federica Valeriani, Antonella Agodi, Beatrice Casini, Maria Luisa Cristina, Marcello Mario D'Errico, Gianluca Gianfranceschi, Giorgio Liguori, Renato Liguori, Nicolina Mucci, Ida Mura, Cesira Pasquarella, Andrea Piana, Giovanni Sotgiu, Gaetano Privitera, Carmela Protano, Annalisa Quattrocchi, Giancarlo Ripabelli, Angelo Rossini, Anna Maria Spagnolo, Manuela Tamburro, Stefano Tardivo, Licia Veronesi, Matteo Vitali, Vincenzo Romano Spica
BackgroundReprocessing of endoscopes is key to preventing cross-infection after colonoscopy. Culture-based methods are recommended for monitoring, but alternative and rapid approaches are needed to improve surveillance and reduce turnover times. A molecular strategy based on detection of residual traces from gut microbiota was developed and tested using a multicenter survey.MethodsA simplified sampling and DNA extraction protocol using nylon-tipped flocked swabs was optimized. A multiplex real-time polymerase chain reaction (PCR) test was developed that targeted 6 bacteria genes that were amplified in 3 mixes. The method was validated by interlaboratory tests involving 5 reference laboratories. Colonoscopy devices (n = 111) were sampled in 10 Italian hospitals. Culture-based microbiology and metagenomic tests were performed to verify PCR data.ResultsThe sampling method was easily applied in all 10 endoscopy units and the optimized DNA extraction and amplification protocol was successfully performed by all of the involved laboratories. This PCR-based method allowed identification of both contaminated (n = 59) and fully reprocessed endoscopes (n = 52) with high sensibility (98%) and specificity (98%), within 3-4 hours, in contrast to the 24-72 hours needed for a classic microbiology test. Results were confirmed by next-generation sequencing and classic microbiology.ConclusionsA novel approach for monitoring reprocessing of colonoscopy devices was developed and successfully applied in a multicenter survey. The general principle of tracing biological fluids through microflora DNA amplification was successfully applied and may represent a promising approach for hospital hygiene.



http://ift.tt/2yDlw4d