PNA is an artificial analog of DNA which the negatively charged phosphodiester backbone is replaced by a neutrally charged N-(2-aminoehtyl)-glycine backbone. To solve those challenges, we developed a Peptide Nucleic Acid based Real Time-LAMP (PNA RT-LAMP) assay “AQ-TOP COVID-19 rapid Detection Kit Plus” which used dual labeled PNA probe that has been reported having superior specificity 19 and sensitivity 20 comparing to the accumulative dye such as SYBR green or sequence specific DNA fluorescent probes which are routinely used in LAMP and other molecular assays. Notably, SARS-CoV-2 LAMP tests have shown inadequate detection performances when testing low positive samples which exhibited Ct values over 30 on real-time PCR assays 14, 15, 16 consequently, LAMP has been considered that having similar diagnostic accuracy as real-time PCR in detection of SARS-CoV-2 in the acute symptomatic phase of COVID-19 while the sample contains high viral load but not in early or late/clearance stages of the infection 17, 18. However, quite a few LAMP tests have been commercialized successfully and multiple scientific reports have shown that the LAMP tests have high enough analytical and clinical performances 6, 7, 8, numerous reports have been showing that the LAMP assays still have some challenges for instance, poor specificity 9, 10, difficulties of establishing multiplexed testing 11, 12, and result interpretation through colorimetric/visual inspection which might be affected by technicians’ subjectivity 13. Since beginning of the COVID-19 outbreak, various molecular detection assays based on real-time PCR or LAMP aimed for detection of SARS-CoV-2 nucleic acid have been developed and received Emergency Use Authorizations (EUA) from the US FDA or the authorizations of the regulatory agencies in the country of production. Loop Mediated Isothermal Amplification (LAMP) is a molecular technique capable of detecting nucleic acids with high sensitivity within a reduced time compared to classical real-time PCR and has been used widely for the detection of viral infections in a time-effective manner 5. Highly specific real-time reverse transcriptase PCR assays have been used for the identification of the SARS-CoV-2 RNA globally however, it requires laboratory-based PCR instruments and needs about 2 h of run-time as well as additional incubation of 15 to 30 min for cDNA synthesis from template RNA 4. The high transmissibility demands rapid and accurate detection of SARS-CoV-2 at the early stages of the infection in a cost-effective manner 3. Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is a single-stranded RNA virus that causes Coronavirus Disease 2019 (COVID-19) which has been spreading globally at rapid speed and is more contagious than most of other human respiratory tract infectious microorganisms 1, 2.
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