Detection of Enterovirus on Environmental Surfaces in Daycare Centers Using Droplet Digital PCR (ddPCR) and Its Public Health Implications

Enteroviruses (EVs) are major pathogens that can be transmitted by direct and indirect contact through respiratory droplets from infected individuals and children are particularly susceptible. EVs survive on surfaces for extended periods; therefore, environmental hygiene is critical in communal environments. We investigated EV presence on environmental surfaces in daycare centers from April to July 2024. Environmental samples (300) were collected from floors, toys, and desks. Viral RNA was extracted and analyzed using real-time reverse transcription polymerase chain reaction (real-time RT-PCR) and ddPCR to detect pan-enterovirus (pan-EVs) and EV D68. Enteroviruses were detected in 45.3% of the samples. Specifically, pan-EV was found in 88 samples (1.12–505 copies/20 μL) and EV D68 in 104 samples (1.12–309 copies/20 μL). The most frequently contaminated surfaces were floors (31%), toys (13%), and desks (11%). Monthly analysis showed a gradual decrease in viral detection rates from 88.6% in April to 18.5% in July, indicating effectiveness of environmental disinfection. Additionally, ddPCR demonstrated nearly twice the detection rate of real-time RT-PCR, achieving high sensitivity. Detection of EVs on various surfaces in daycare centers, highlights their potential for indirect transmission. Thus, systematic environmental disinfection protocols and systematic hygiene management must be implemented to prevent EV spread.