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.

Potential testing of reprocessing procedures by real-time polymerase chain reaction: A multicenter study of colonoscopy devices / Valeriani, F.; Agodi, A.; Casini, B.; Cristina, M. L.; D'Errico, M. M.; Gianfranceschi, G.; Liguori, G.; Liguori, R.; Mucci, N.; Mura, I.; Pasquarella, Cesira Isabella Maria; Piana, A.; Sotgiu, G.; Privitera, G.; Protano, C.; Quattrocchi, A.; Ripabelli, G.; Rossini, A.; Spagnolo, A. M.; Tamburro, M.; Tardivo, S.; Veronesi, Licia; Vitali, M.; Romano, Spica V.. - In: AMERICAN JOURNAL OF INFECTION CONTROL. - ISSN 0196-6553. - 46:2(2017), pp. 159-164. [10.1016/j.ajic.2017.08.008]

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

PASQUARELLA, Cesira Isabella Maria;VERONESI, Licia;
2017-01-01

Abstract

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.
Potential testing of reprocessing procedures by real-time polymerase chain reaction: A multicenter study of colonoscopy devices / Valeriani, F.; Agodi, A.; Casini, B.; Cristina, M. L.; D'Errico, M. M.; Gianfranceschi, G.; Liguori, G.; Liguori, R.; Mucci, N.; Mura, I.; Pasquarella, Cesira Isabella Maria; Piana, A.; Sotgiu, G.; Privitera, G.; Protano, C.; Quattrocchi, A.; Ripabelli, G.; Rossini, A.; Spagnolo, A. M.; Tamburro, M.; Tardivo, S.; Veronesi, Licia; Vitali, M.; Romano, Spica V.. - In: AMERICAN JOURNAL OF INFECTION CONTROL. - ISSN 0196-6553. - 46:2(2017), pp. 159-164. [10.1016/j.ajic.2017.08.008]
File in questo prodotto:
File Dimensione Formato  
1-s2.0-S0196655317309550-main.pdf

accesso aperto

Descrizione: Articolo principale
Tipologia: Versione (PDF) editoriale
Licenza: Creative commons
Dimensione 289.29 kB
Formato Adobe PDF
289.29 kB Adobe PDF Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11381/2832903
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 21
  • ???jsp.display-item.citation.isi??? 18
social impact