Rapid Detection of SARS-CoV-2 Using Reverse transcription RT-LAMP method

Abstract

Corona Virus Disease 2019 (COVID-19) is a recently emerged life-threatening disease caused by SARS-CoV-2. Real- time fluorescent PCR (RT-PCR) is the clinical standard for SARS-CoV-2 nucleic acid detection. To detect SARS-CoV-2 early and control the disease spreading on time, a faster and more convenient method for SARS-CoV-2 nucleic acid detecting, RT-LAMP method (reverse transcription loop-mediated isothermal amplification) was developed. RNA reverse transcription and nucleic acid amplification were performed in one step at 63 ℃ isothermal conditions, and the results can be obtained within 30 minutes. ORF1ab gene, E gene and N gene were detected at the same time. ORF1ab gene was very specific and N gene was very sensitivity, so they can guarantee both sensitivity and specificity for SARS-CoV-2. The sensitivity of RT-LAMP assay is similar to RT-PCR, and specificity was 99% as detecting 208 clinical specimens. The RT-LAMP assay reported here has the advantages of rapid amplification, simple operation, and easy detection, which is useful for the rapid and reliable clinical diagnosis of SARS-CoV-2.

COVID‐19 diagnostic testing: Technology perspective

“…38 Similar results were obtained in an independent study with 17 and 191 patients who had or did not have COVID-19, respectively. 40 A major drawback of RT-LAMP is that it is low throughput because only one sample can be measured in an experiment.…”

LAMP-Seq: Population-Scale COVID-19 Diagnostics Using Combinatorial Barcoding

“…RT-LAMP reactions have been demonstrated to be highly sensitive for sequence-specific viral nucleic acid detection (Lamb et al, 2020;Yang et al, 2020;Zhang et al, 2020), even from unpurified samples (Estrela et al, 2019). To establish a barcoded RT-LAMP reaction, we inserted barcode sequences into the forward inner primer (FIP), which enables generation of repeatedly barcoded palindromic amplification products ( Fig.…”

Development of a dual-gene loop-mediated isothermal amplification (LAMP) detection assay for SARS-CoV-2: A preliminary study

“…In summary, it is well-established that LAMP is a stable and rapidly deployable approach with sensitivity profile surpassing the PCR [10,14,15]. Recently, several LAMP assays for the detection of SARS-CoV-2 have been reported and showed promising data [16][17][18][19][20][21][22][23]. In the present study, we report a novel SARS-CoV-2 dual gene RT-LAMP assay and its preliminary evaluation on "real-world" cases.…”