Enhancing efficiency in detection of COVID-19 through AI-driven colorimetric isothermal detection with multiplex primers

• Published in: Diagnostic microbiology and infectious disease 2024-10, Vol.110 (2), p.116446, Article 116446
• Main Authors: Jaroenram, Wansadaj, Teerapittayanon, Surat, Suvannakad, Rapheephat, Pengpanich, Sukanya, Kampeera, Jantana, Arunrut, Narong, Dangtip, Sirintip, Sirithammajak, Sarawut, Tondee, Benyatip, Khumwan, Pakapreud, Japakasetr, Supak, Leaungwutiwong, Pornsawan, Chatnuntawech, Itthi, Kiatpathomchai, Wansika
• Format: Article
• Language: English
• Subjects: Colorimetric RT-LAMP; Colorimetry - methods; COVID-19 - diagnosis; COVID-19 - virology; COVID-19 Nucleic Acid Testing - methods; COVID-19 Testing - methods; Deep learning; DNA Primers - genetics; Humans; Machine learning; Molecular Diagnostic Techniques - methods; Molecular Diagnostic Techniques - standards; Multiplex RT-LAMP; Nucleic Acid Amplification Techniques - methods; Rapid detection; SARS-CoV-2; SARS-CoV-2 - genetics; SARS-CoV-2 - isolation & purification; Sensitivity and Specificity; Smartphone
• ISSN: 0732-8893; 1879-0070; 1879-0070
• DOI: 10.1016/j.diagmicrobio.2024.116446

•The strategy of using multiplex primers to enhance colorimetric LAMP assay's efficiency was demonstrated with success.•The multiplex primers enhanced the sensitivity of the existing RT-LAMP assay by 10–100 times.•The proposed assay can detect variant of concerns (VoCs) of COVID-19.•The AI-assisted result analysis can analyze approximately 6700 samples/minute, thereby demonstrating its efficiency and scalability in point-of-care screening. COVID-19 has afflicted millions of lives worldwide. Although there are many rapid methods to detect it based on colorimetric loop-mediated isothermal amplification, there remains room for improvement. This study aims to 1) integrate multiple primers into a singleplex assay to enhance the diagnostic sensitivity, and 2) utilize a high-throughput smartphone-operatable AI-driven color reading tool to enable a rapid result analysis. This setup can improve the sensitivity by 10–100 times and can analyze approximately 6700 samples per minute. The assay is simpler than RT-qPCR, with a turnaround time of less than 75 min. It can detect various types of SARS-CoV-2 by targeting 3 genes, increasing the likelihood that it will remain effective even if the virus undergoes mutations in any single target gene. In summary, it affords potential for adaptation to detection of new/re-emerging diseases with the visual readout for maximum assay simplicity and AI-operated mode for large-scale testing.