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Abstract
ABSTRACTHands play a critical role in the transmission of microbiota on one’s own body, between individuals, and on environmental surfaces. Effectively measuring the composition of the hand microbiome is important to hand hygiene science, which has implications for human health. Hand hygiene products are evaluated using standard culture-based methods, but standard test methods for culture-independent microbiome characterization are lacking. We sampled the hands of 50 participants using swab-based and glove-based methods prior to and following four hand hygiene treatments (using a nonantimicrobial hand wash, alcohol-based hand sanitizer [ABHS], a 70% ethanol solution, or tap water). We compared results among culture plate counts, 16S rRNA gene sequencing of DNA extracted directly from hands, and sequencing of DNA extracted from culture plates. Glove-based sampling yielded higher numbers of unique operational taxonomic units (OTUs) but had less diversity in bacterial community composition than swab-based sampling. We detected treatment-induced changes in diversity only by using swab-based samples (P < 0.001); we were unable to detect changes with glove-based samples. Bacterial cell counts significantly decreased with use of the ABHS (P < 0.05) and ethanol control (P < 0.05). Skin hydration at baseline correlated with bacterial abundances, bacterial community composition, pH, and redness across subjects. The importance of the method choice was substantial. These findings are important to ensure improvement of hand hygiene industry methods and for future hand microbiome studies. On the basis of our results and previously published studies, we propose recommendations for best practices in hand microbiome research.
Highlights
IMPORTANCE The hand microbiome is a critical area of research for diverse fields, such as public health and forensics
The effects of hand hygiene practices on the skin and hand microbiome are an important area of study, but relatively few robust studies have been conducted to fully elucidate the optimal protocols to ensure repeatability, precision, and accuracy
Wellestablished protocols are currently available from the hand hygiene industry to assess
Summary
IMPORTANCE The hand microbiome is a critical area of research for diverse fields, such as public health and forensics. The use of 16S rRNA gene sequencing methods has become routine in microbiology studies For these reasons, cultureindependent methods are expected to continue to be used more frequently in many fields, including hand hygiene effect research and product development. In the techniques in the second category, which applies to surgical scrubs (e.g., ASTM E1115 [18]), the objective is to evaluate the hand hygiene product for its ability to reduce resident bacteria on hands In both categories, hands are sampled using the “glove-based” methodology by vigorous massage in loose-fitting gloves with an eluent for 1 min, after which the eluents are assayed for bacteria using culture-based methods [10].
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