TY - JOUR T1 - Development Optimization and Validation of RT-LAMP based COVID-19 Facility in Pakistan JF - bioRxiv DO - 10.1101/2020.05.29.124123 SP - 2020.05.29.124123 AU - Farhan Haq AU - Salmaan Sharif AU - Adnan Khurshid AU - Imran Shabbir AU - Muhammad Salman AU - Nazish Badar AU - Aamir Ikram AU - Abdul Ahad AU - Muhammad Faraz Arshad Malik Y1 - 2020/01/01 UR - http://biorxiv.org/content/early/2020/05/29/2020.05.29.124123.abstract N2 - The pandemic SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2) has created a widespread panic across the globe especially in the developing countries like Pakistan. The lack of resources and technical staff are causing havoc challenges in the detection and prevention of this global outbreak. Therefore, a less expensive and massive screening of suspected individuals for COVID-19 is required. In this study, a user-friendly technique of reverse transcription-loop mediated isothermal amplification (RT-LAMP) was designed and validated to suggest a potential RT-qPCR alternate for rapid testing of COVID-19 suspected individuals. A total of 12 COVID-19 negative and 72 COVID-19 suspected individuals were analyzed. Both RT-qPCR and RT-LAMP assays were performed for all the individuals using open reading frame (ORF 1ab), nucleoprotein (N) and Spike (S) genes. All 12 specimens which were negative using RT-qPCR were also found negative using RT-LAMP assay. Overall 62 out of 72 positive samples (detected using RT-qPCR) were found COVID-19 positive using RT-LAMP assay. Interestingly all samples (45) having Ct values less than 30 showed 100% sensitivity. However, samples with weaker Ct values (i.e., => 35) showed 54% concordance, suggesting potential false negatives or false positives in RT-LAMP or RT-qPCR results, respectively.Overall comparative assessment showed that RT-LAMP assay showed strong sensitivity and specificity and can be used as an alternative strategy for rapid COVID-19 testing. Hence, based on fast processing time, minimal risk of specimens transfer and utilizing available resources, LAMP based detection of COVID-19 is strongly advocated especially for developing countries.Competing Interest StatementThe authors have declared no competing interest. ER -