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Performance and functional assessment of the Kimera P-IV point-of-care plasmonic qPCR prototype for ultra rapid pathogen detection of chlamydia trachomatis

Authors: Hayes JLee SSCarnevale JShamir DBohbot MKirk AGPaliouras MTrifiro MA


Affiliations

1 Lady Davis Institute for Medical for Medical Research - Jewish General Hospital, Montreal, QC, Canada.
2 Division of Experimental Medicine, McGill University, Montreal, QC, Canada.
3 Department of Biology, Concordia University, Montreal, QC, Canada.
4 Nexless Healthcare LP, Montreal, QC, Canada.
5 Department of Electrical and Computer Engineering, McGill University, Montreal, QC, Canada.
6 Department of Medicine, McGill University, Montreal, QC, Canada.

Description

Current standard microbiological techniques are generally very time consuming, usually requiring 24-72 h to establish a diagnosis. Consequentially, contemporary clinical practices implement broad-spectrum antibiotic administration prior to pathogen detection, prompting the emergence of extremely dangerous antibiotic-resistant bacteria. Additionally, lengthy test-to-result turnover times can greatly exacerbate the rate of disease spread. Rapid point-of-care (POC) diagnostics has quickly gained importance since the SARS-CoV-2 pandemic; accordingly, we have developed a rapid four-channel POC plasmonic quantitative polymerase chain reaction (qPCR) machine (Kimera P-IV) to respond to the deficiencies in infection control. Utilizing gold nanorods (GNRs) as nano-heaters and integrating vertical cavity surface emitting lasers (VCSEL) to replace traditional Peltier blocks, the Kimera P-IV has also incorporated quantitative real-time fluorescent monitoring. Using Chlamydia trachomatis genetic material to evaluate the rapid thermocycling performance of the platform, we have generated positive amplicons in less than 13 min; however, to achieve these results, several biological reagent considerations needed to be taken into account, specifically primer design. The device can achieve a limit of detection (LoD) of <101 DNA copies, a PCR efficiency of 88.3%, and can differentiate positive from negative results with 100% accuracy. Moreover, it can also analyze C. trachomatis DNA spiked urine samples via a simple dilution, suggesting that a separate nucleic acid step may not be needed for diagnosing infections. In conclusion, the operation of the Kimera P-IV prototype places it in a unique position of POC devices to revolutionize infectious disease diagnosis.


Links

PubMed: https://pubmed.ncbi.nlm.nih.gov/39881625/

DOI: 10.1017/S0950268825000081