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Background <p>Public washrooms (toilets) are potential hubs for pathogen transmission, yet the risk of microbial re-contamination via post-handwashing surfaces remains understudied. We characterized the prevalence and distribution of multidrug-resistant organisms (MDROs) and antimicrobial resistance genes (ARGs) in post-handwashing areas by sampling four high-contact sites, including faucets, paper dispensers, hand dryers, and exit door handles, in public washrooms across healthcare, commercial, and recreational facilities.</p> Results <p>From the 232 post-handwashing surface samples collected, we isolated 17 MDROs (7.33% prevalence) from cultures, including extended-spectrum beta-lactamase (ESBL)-producing <i>Enterobacterales</i> (ESBL-E, <i>n</i> = 10), carbapenem-resistant <i>Pseudomonas aeruginosa</i> (CRPA, <i>n</i> = 5), and methicillin-resistant <i>Staphylococcus aureus</i> (MRSA, <i>n</i> = 2). Additionally, we novelly employed targeted probe capture metagenomics (TCM), which utilizes oligonucleotide probes to enrich and detect low-abundance microbial species and ARG sequences. TCM revealed the detection of human pathogenic taxa in 65.2% of samples, including <i>P. aeruginosa</i> (78.4%), <i>Acinetobacter baumannii</i> (77.9%), and <i>S. aureus</i> (71.1%). Clinically critical ARGs, such as bla<i>CTX-M</i> (2.0%), bla<i>NDM</i> (2.9%), bla<i>SHV</i> (3.4%), and <i>mecA</i> (62.3%), were detected in 63.7% of samples, indicating a potential transmission within the post-handwashing area.</p> Conclusions <p>Our findings highlight the role of post-handwashing areas as underrecognized reservoirs for MDROs, particularly MRSA. Furthermore, this study demonstrates the utility of TCM in public health surveillance by enabling a sensitive detection of rare but high-risk microbial species and drug resistance determinants in low-biomass environmental samples. This study offers a comprehensive and nuanced view of the microbial and resistome landscape of washroom environments, offering a revolutionary approach for future environmental surveillance.</p>

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Targeted probe capture metagenomics-enabled surveillance of multidrug-resistant organisms and antimicrobial resistance genes in post-handwashing areas of public washrooms

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年11月17日

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  • Alison Yee-Ting Lam,
  • Chun-Hei Lau,
  • Wing-Yin Tam,
  • Chloe Toi-Mei Chan,
  • Tsun-Ming Lok,
  • Lorna Kwai-Ping Suen,
  • Lam-Kwong Lee,
  • Elaine Yin-Ying Yeung,
  • Tsz-Kei Lam,
  • Wai-Kam Cheung,
  • Man-Wa Chui,
  • Ho-Sing Soong,
  • Franklin Wang-Ngai Chow,
  • Simon Ching Lam,
  • Sony Nai-Yeung So,
  • Sam Kit-San Yuen,
  • Gilman Kit-Hang Siu

摘要

Background

Public washrooms (toilets) are potential hubs for pathogen transmission, yet the risk of microbial re-contamination via post-handwashing surfaces remains understudied. We characterized the prevalence and distribution of multidrug-resistant organisms (MDROs) and antimicrobial resistance genes (ARGs) in post-handwashing areas by sampling four high-contact sites, including faucets, paper dispensers, hand dryers, and exit door handles, in public washrooms across healthcare, commercial, and recreational facilities.

Results

From the 232 post-handwashing surface samples collected, we isolated 17 MDROs (7.33% prevalence) from cultures, including extended-spectrum beta-lactamase (ESBL)-producing Enterobacterales (ESBL-E, n = 10), carbapenem-resistant Pseudomonas aeruginosa (CRPA, n = 5), and methicillin-resistant Staphylococcus aureus (MRSA, n = 2). Additionally, we novelly employed targeted probe capture metagenomics (TCM), which utilizes oligonucleotide probes to enrich and detect low-abundance microbial species and ARG sequences. TCM revealed the detection of human pathogenic taxa in 65.2% of samples, including P. aeruginosa (78.4%), Acinetobacter baumannii (77.9%), and S. aureus (71.1%). Clinically critical ARGs, such as blaCTX-M (2.0%), blaNDM (2.9%), blaSHV (3.4%), and mecA (62.3%), were detected in 63.7% of samples, indicating a potential transmission within the post-handwashing area.

Conclusions

Our findings highlight the role of post-handwashing areas as underrecognized reservoirs for MDROs, particularly MRSA. Furthermore, this study demonstrates the utility of TCM in public health surveillance by enabling a sensitive detection of rare but high-risk microbial species and drug resistance determinants in low-biomass environmental samples. This study offers a comprehensive and nuanced view of the microbial and resistome landscape of washroom environments, offering a revolutionary approach for future environmental surveillance.