Detection of biological loads in sewage using the automated robot-driven
photoelectrochemical biosensing platform
Abstract
Real-time polymerase chain reaction (RT-PCR) remains the most prevalent
molecular detection technology for sewage analysis but is plagued with
numerous disadvantages, such as time consumption, stringent equipment
requirements, and susceptibility to false negatives. In this study, we
construct an automated robot-driven photoelectrochemical biosensing
platform that utilizes the CRISPR/Cas12a system, to achieve fast,
ultrasensitive, high specificity detection of biological loads in
sewage. The Shennong-1 robot integrates several functional modules,
involving sewage sampling and pretreatment to streamline the sewage
monitoring. A screen-printed electrode is employed with a vertical
graphene-based working electrode and enhanced with surface-deposited Au
nanoparticles (NPs). CdTe/ZnS quantum dots (QDs) are further fabricated
through the double-stranded DNA anchored on Au nanoparticles. Using cDNA
template of Omicron BA.5 spike gene as a model, the photoelectrochemical
biosensor demonstrates excellent analytical performance, with a lower
detection limit of 2.93×102 zM and an outstanding selectivity at the
level of single-base mutation recognition. Furthermore, the rapid,
accurate detection of BA.5 in sewage demonstrates the feasibility of the
photoelectrochemical platform for sewage monitoring. In conclusion, this
platform allows early detection and tracking of infectious disease
outbreaks, providing timely data support for public health institutions
to take appropriate prevention and control measures.