Rapid Quantification of the Total Viable Bacterial Population on Human Hands Using Flow Cytometry with SYBR® Green I

Background: Monitoring the efficiency of hand washing typically relies on determining the presence or absence of targeted organisms on the hands, not necessarily determining if the method is working. The results focus on the organism studied and do not give a true reflection of the efficiency of the hygiene intervention. To obtain a better picture, this study tested flow cytometry with SYBR® Green I to quantify bacterial populations on the hands of participants in a healthcare training clinic at a University in South Africa. Methods: Participants “washed” both hands in a buffer solution in a sterile bag, and the total and viable bacterial populations numbers were determined using flow cytometry and compared with standard culture-based methods, testing for total coliforms, E. coli and heterotrophic organisms (IDEXX Colilert®-18 Quanti-Trays™ and SimPlates®) and fastidious organisms with hemolytic activity (Blood Agar). Results: Compared to the culture-based method, flow cytometry rapidly distinguished total, viable, and dead bacterial populations in all samples. As expected, the culture-based methods gave lower bacterial counts and were limited by the detection limit of each test, requiring further testing with additional costs. Although the exact bacterial species in the population could not be identified with flow cytometry, viable counts from flow cytometric analysis were two times more precise than the data obtained with any of the culture-based methods tested. Conclusions: Flow cytometry was found to be suitable for immediate, accurate, and automatic detection of bacteria, and because it describes the viable bacterial population, it is ideal to monitor hand hygiene interventions.