Control of the efficiency of instrument-cleaning using real-time Polymerase Chain Reaction

• Published in: Zentralblatt für Chirurgie - Zeitschrift für Allgemeine, Viszeral-, Thorax- und Gefäßchirurgie
• Main Authors: Stoleriu, MG, Avci-Adali, M, Wendel, HP, Schlensak, C, Walker, T
• Format: Conference Proceeding
• Language: English; German
• Published: 09.09.2015
• ISSN: 0044-409X; 1438-9592
• DOI: 10.1055/s-0035-1559923

Background: Inefficiently cleaned surgical instruments may cause the transfer of infections. The control of the efficieny of medical cleaning of reusable instruments is a challenge in every medical facility. Various control-methods have been developed over time. Here we present the use of the Polymerase Chain Reaction (PCR) to evaluate the success of cleaning efforts of surgical instruments. Methods: Stainless steel surgical instruments were “contaminated” with defined synthetic Single-Stranded DNA (ssDNA) – containing solutions. After undergoing the cleaning process rests of the ssDNA were washed out of the instruments using solutions that do not interfere with the effectiveness of the polymerase. The residual ssDNA- amount in these eluates was detected using real-time PCR. Results: Through the use of sequence specific ssDNA samples and real-time PCR we were able to detect and quantify the residual DNA amount on the instruments. Since we used defined ssDNA- specific primers we could avoid the amplification of possible residual genomic DNA on the instruments e.g. through pollution with patient-DNA material. As our defined ssDNA cannot be found in nature we could thus achieve a ultra high sensitivity in our testing method. Discussion: Quantitative PCR methods are a common technique to both detect and quantify DNA samples. Here we used this method in order to detect ssDNA samples on surgical instruments after medical cleaning. This control method appears to be highly sensitive as well as specific and should be considered a valuable instrument to evaluate cleaning methods of reusable instruments in order to reduce undesirable infections.