RT-LAMP assay for ultra-sensitive detection of SARS-CoV-2 in comparison with RT-qPCR

Seyed Mohammad Ghorbani Dazmiri¹ Seyyed Danial Mirmiran² Fatemeh Sadat Abolhasani³ Elmira Karimonafs⁴ Parivash Ghaderinia⁵ Shila Fallahpour⁶ Hooman Hanifehpour⁷

1) Faculty of Biological Sciences, Sari Branch, Islamic Azad University, Sari, Iran
2) State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University Wuhan, China
3) Department of Microbiology, Faculty of Biological Sciences, Alzahra University, Tehran ,Iran
4) Department of Microbiology, Faculty of Biological Sciences, North Tehran Branch, Islamic Azad University, Tehran, Iran
5) Department of Microbiology, Zanjan Branch, Islamic Azad University, Zanjan, Iran
6) Department of Biology, Biological Science Faculty, Islamic Azad University of Falavarjan, Isfahan, Iran
7) Department of Microbiology & Immunology, Louis Pasteur University, Strasburg, France

Publication : 2nd International Conference on Recent Advances in Engineering, Innovation and Technology - Belgium(eitconf.com)
Abstract :
To combat the COVID-19 pandemic, it is critical to quickly and accurately detect SARS-CoV-2 infection in patients since there are currently no effective antiviral drugs or vaccines available. This study aimed to create and evaluate a rapid One-Step LAMP assay that can directly detect SARS-CoV-2 RNA from nasopharyngeal swab samples of patients suspected of having the infection in comparison with RT-qPCR assay. In this research, a cross-sectional survey involved testing 225 nasopharyngeal swab samples from patients suspected of having COVID-19 using two different methods: One-Step RT-qPCR and One-Step LAMP assay. The One-Step LAMP assay directly amplified the extracted RNA from the SARS-CoV-2 nasopharyngeal sample using a new version of the key enzyme of the LAMP reaction called Bst DNA/RNA Polymerase 3.0 (Bst 3). The study evaluated the accuracy and reliability of the One-Step LAMP assay by comparing it with One-Step RT-qPCR using positive and negative clinical samples of COVID-19. Out of the 225 participants, the One-Step RT-qPCR and One-Step LAMP assays detected positive SARS-CoV-2 results in 115(51.1%) and 110(48.8%) participants, respectively. The Cohen s kappa coefficient (κ) demonstrated that there was a 97% agreement rate between the two tests, which was statistically significant (P <0.001), The study found a significant relationship (P <0.001) between the variables of residency, clinical signs of sore throat, fever and chills, shortness of breath, underlying health conditions (UHC), LICT, and infection with SARS-CoV-2. The study concluded that the One-Step LAMP assay is an effective method for detecting SARS-CoV-2 in suspected patients compared to the One-Step RT-qPCR method due to its low cost, simplicity, sensitivity, and specificity. Therefore, the One-Step LAMP assay has significant potential as a valuable diagnostic tool for disease control, timely treatment, and protecting public health during epidemics.
Keywords : Rapid Test sensitive SARS-CoV-2 One-Step LAMP One-Step RT-qPCR