ATP bioluminescence assay for evaluating cleaning practices in operating theatres: applicability and limitations

Abstract

BackgroundEnvironmental cleaning practice plays an important role in reducing microbial contamination in hospital surfaces and contributes to prevent Healthcare Associated Infections. Adenosine Triphosphate (ATP) bioluminescence assay is a commonly used method for assessing environmental cleanliness on healthcare surfaces. This study tested the feasibility of using ATP-bioluminescence assay for evaluating the efficiency of cleaning procedures in the operating theatre settings, comparing the ATP-bioluminescence test with the traditional culture method.MethodsThe surfaces of 10 operating rooms of two public hospitals (140 samples in total) were examined “at rest”, in two moments of the same daily session: before the first scheduled operation (Pre), and before the second, after a clean environment was re-established (Post). Surface contamination was assessed using the cultural method to detect Total Viable Counts (TVC36°C) and ATP-bioluminescence assay (RLU).ResultsThe examined surfaces presented very low TVCs (geometric means: 1.8 CFU/plate; IC95%: 1.6–2.0), always compliant with the relative reference standards. No statistical correlation was found between ATP values and TVCs. However, considering the results in terms of general evaluation of hygienic quality of surfaces, the two methods were consistent in identifying the most contaminated areas (Hospital A > Hospital B; Pre > Post; most contaminated surfaces: scialytic lamp). Furthermore, the ATP mean values showed a progressive increase from surfaces with TVC = 0 to surfaces with TVC > 15 CFU/plate.ConclusionsAlthough not an alternative to cultural methods, the ATP-bioluminescence-assay can be a useful tool to measure the efficiency of cleaning procedures also in environments with very low microbial counts. Each health facility should identify appropriate reference values, depending on the devices used and on the basis of the analysis of the data collected through spatial and temporal sampling series. By providing a rapid feedback, the ATP-assay helps to increase the awareness of operators and allows immediate action to be taken in critical situations.

Evaluation of the complexity of indoor air in hospital wards based on PM2.5, real-time PCR, adenosine triphosphate bioluminescence assay, microbial culture and mass spectrometry

“…The traditionally applied methods, to assess microbial air contamination and cleanliness on surfaces in hospital are based on microbiological cultural technology, including active, passive and surfaces sampling [27]. In recent decade, adenosine triphosphate (ATP) bioluminescence assay, which was used to assess environmental surface cleanliness has been proposed [28]. Previous studies have shown that the number of microorganisms that can be cultured only accounts for < 5% of the total microorganisms in air [29–31].…”

The Influence of Surface Topography and Wettability on Escherichia coli Removal from Polymeric Materials in the Presence of a Blood Conditioning Film

“…This indicated that the ATP quantified prior to the sanitisation process did not correlate with the cell counts observed. The quantification of ATP is an indicator of organic material and is not exclusive to microbial contamination, and whilst studies have shown a correlation between ATP levels and total viable counts, a direct relationship remains unclear [ 56 , 57 , 58 ].…”

“…Limitations of this study include bacterial enumeration via CFU/mL, since this method does not account for viable but non-culturable (VBNC) cells [ 71 ]. Finally, although ATP quantification is easy to use and rapid, it is an indicator for organic material and not solely microbial contamination [ 56 , 57 , 58 ].…”

Application of Biosensors for Detection of Pathogenic Food Bacteria: A Review

“…The bioluminescent sensors of whole cells have benefits over conventional approaches by being faster, more cost effective, easy to carry out and less labor-intensive [198]. While not an alternative to traditional approaches, an ATP-bioluminescence-assay can also be a valuable tool for determining the efficacy of environmental cleanliness procedures even with very low microbial counts [199]. Moreover, bioluminescent techniques often possess several benefits compared to fluorometric techniques mainly because no wavelength of excitation is required for the representation of light.…”