A new diagnostic test can detect the RNA of SARS-CoV-2, the virus that causes COVID-19, in urine, blood, saliva or mouth swab samples in just 30 to 45 minutes, according to a study by Laura Lamb of the Beaumont Health System, Michigan, US, and colleagues.
SARS-CoV-2 is difficult to diagnose early in infection as patients may be asymptomatic or have mild, non-specific symptoms. The current standard for diagnostic molecular is quantitative reverse transcription PCR (qRT-PCR), which requires expensive equipment and trained personnel only available in limited laboratories. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a one-step technology with fewer barriers to use than qRT-PCR; reagents are inexpensive and can be stored at room temperature, the technique works with a range of sample types, and it has been found to be highly specific, sensitive, and fast for other infectious diseases.
In the new study, researchers developed an RT-LAMP diagnostic test for SARS-CoV-2 and measured the effectiveness of the test on blood, urine, saliva and mouth swab samples that had been spiked with different amounts of SARS-CoV-2 genetic material. The test was also used on clinical samples from COVID-19 patients.
The RT-LAMP test for SARS-CoV-2 successfully detected the virus in all human sample types that were spiked with SARS-CoV-2 genetic material, within 30 to 45 minutes, with an estimated limit of detection of as few as 304 viral copies in a sample. Importantly, the test did not detect virus in samples spiked with RNA other coronaviruses, demonstrating specificity for SARS-CoV-2. In clinical mouth swab samples, RT-LAMP was positive for 95% (19/20) of samples positive by qRT-PCR and negative for 90% (18/20) of samples that were negative by qRT-PCR. The researchers note that those discrepancies between the methods could represent either false positives by the RT-LAMP, contamination or increased sensitivity of RT-LAMP compared to qRT-PCR. The current study was not powered to determine sensitivity in a clinical population.
Lamb summarises: “This method can rapidly detect the virus for COVID-19 in clinical samples without expensive equipment.”
Abstract
Novel Corona virus/Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2 or 2019-nCoV), and the subsequent disease caused by the virus (coronavirus disease 2019 or COVID-19), is an emerging global health concern that requires a rapid diagnostic test. Quantitative reverse transcription PCR (qRT-PCR) is currently the standard for SARS-CoV-2 detection; however, Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) may allow for faster and cheaper field based testing at point-of-risk. The objective of this study was to develop a rapid screening diagnostic test that could be completed in 30–45 minutes. Simulated patient samples were generated by spiking serum, urine, saliva, oropharyngeal swabs, and nasopharyngeal swabs with a portion of the SARS-CoV-2 nucleic sequence. RNA isolated from nasopharyngeal swabs collected from actual COVID-19 patients was also tested. The samples were tested using RT-LAMP as well as by conventional qRT-PCR. Specificity of the RT-LAMP was evaluated by also testing against other related coronaviruses. RT-LAMP specifically detected SARS-CoV-2 in both simulated patient samples and clinical specimens. This test was performed in 30–45 minutes. This approach could be used for monitoring of exposed individuals or potentially aid with screening efforts in the field and potential ports of entry.
Authors
Laura E Lamb, Sarah N Bartolone, Elijah Ward, Michael B Chancellor
[link url="https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0234682"]PLOS One abstract[/link]