Potential testing of reprocessing procedures by real-time polymerase chain reaction: A multicenter study of colonoscopy devices
| Autor: | Valeriani, Federica, Agodi, Antonella, Casini, Beatrice, Cristina, Maria Luisa, D'Errico, Marcello Mario, Gianfranceschi, Gianluca, Liguori, Giorgio, Liguori, Renato, Mucci, Nicolina, Mura, Ida, Pasquarella, Cesira, Piana, Andrea, Sotgiu, Giovanni, Privitera, Gaetano, Protano, Carmela, Quattrocchi, Annalisa, Ripabelli, Giancarlo, Rossini, Angelo, Spagnolo, Anna Maria, Tamburro, Manuela, Tardivo, Stefano, Veronesi, Licia, Vitali, Matteo, Romano Spica, Vincenzo, GISIO Working Group of the Italian Society of Hygiene, Preventive Medicine and Public Health. |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
DNA Bacterial medicine.medical_specialty Quality Assurance Health Care Epidemiology 030106 microbiology Colonoscopy Flocked swab sampling Health care-associated infections MfDNA Sanitation Surveillance Epidemiology Health Policy Public Health Environmental and Occupational Health Infectious Diseases Real-Time Polymerase Chain Reaction law.invention 03 medical and health sciences 0302 clinical medicine law Medicine Humans Multiplex Medical physics Sanitation Polymerase chain reaction Protocol (science) Flocked swab sampling Surveillance medicine.diagnostic_test Colonoscopes business.industry Health Policy Environmental and Occupational Health Public Health Environmental and Occupational Health Microbiology Test Health care-associated infections MfDNA Infectious Diseases mfDNA DNA extraction Surgery Disinfection Real-time polymerase chain reaction Multicenter study flocked swab sampling health care-associated infections mfdna sanitation surveillance epidemiology health policy public health environmental and occupational health infectious diseases Equipment Contamination 030211 gastroenterology & hepatology Public Health business |
| Popis: | Background Reprocessing 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. Methods A 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. Results The 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. Conclusions A 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. |
| Databáze: | OpenAIRE |
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