Considerations for the selection of tests for SARS-CoV-2 molecular diagnostics

Abstract

During the course of 2020, the outbreak of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS‐CoV‐2) spread rapidly across the world. Clinical diagnostic testing for SARS-Cov-2 infection has relied on the real‐time Reverse Transcriptase Polymerase Chain Reaction and is considered the gold standard assay. Commercial vendors and laboratories quickly mobilised to develop diagnostic tests to detect the novel coronavirus, which was fundamentally important in the pandemic response. These SARS-Cov-2 assays were developed in line with the Food Drug Administration-Emergency Use Authorization guidance. Although new tests are continuously being developed, information about SARS-CoV-2 diagnostic molecular test accuracy has been limited and at times controversial. Therefore, the analytical and clinical performance of SARS-CoV-2 test kits should be carefully considered by the appropriate regulatory authorities and evaluated by independent laboratory validation. This would provide improved end-user confidence in selecting the most reliable and accurate diagnostic test. Moreover, it is unclear whether some of these rapidly developed tests have been subjected to rigorous quality control and assurance required under good manufacturing practice. Variable target gene regions selected for currently available tests, potential mutation in target gene regions, non-standardized pre-analytic phase, a lack of manufacturer independent validation data all create difficulties in selecting tests appropriate for different countries and laboratories. Here we provide information on test criteria which are important in the assessment and selection of SARS-CoV-2 molecular diagnostic tests and outline the potential issues associated with a proportion of the tests on the market. Supplementary Information The online version contains supplementary material available at 10.1007/s11033-022-07455-5.

“It Was Very Comforting to Find Out Right Away.” – Patient Perspectives on Point-of-Care Molecular SARS-CoV-2 Testing in Primary Care

“… 5 , 6 However, laboratory testing using the real-time quantitative reverse transcription–polymerase chain reaction (RT-qPCR) technology remains the gold standard in the diagnosis of acute SARS-CoV-2 infection. 7 A positive test result has far-reaching consequences for patients and their social network such as quarantine. The rapid availability of the test result could be of particular importance in the context of a disease like COVID-19, which possesses a significant public health relevance.…”

Highly sensitive and rapid detection of SARS‐CoV‐2 via a portable CRISPR‐Cas13a‐based lateral flow assay

“…Similarly, all RT‐PCR‐negative samples tested negative following our CRISPR immunochromatography method. Although the sample size was inadequate to evaluate the overall validity of our method, the available data indicate that the effectiveness of this CRISPR immunochromatography method was consistent with that of the gold‐standard RT‐PCR method 22 . Notably, the CRISPR immunochromatography method can report testing results within 1 h, thus saving 30 min to 1 h when compared with RT‐PCR.…”

“…The LoD restriction for the commercial pathogenic nucleic acid detection kit in China is no more than 1 copy/μl; our CRISPR‐LFA method demonstrated an LoD of 0.25 copy/μl and was found to be ultrasensitive. Importantly, this LoD, namely the analytical sensitivity, was comparable to that of the RT‐PCR method or even higher than that of many commercial RT‐PCR detection kits 17,22,23 . The improved sensitivity of our method is mainly owing to various optimizations of the whole reaction system.…”

“…Importantly, this LoD, namely the analytical sensitivity, was comparable to that of the RT‐PCR method or even higher than that of many commercial RT‐PCR detection kits. 17 , 22 , 23 The improved sensitivity of our method is mainly owing to various optimizations of the whole reaction system. The sequences of the MIRA primer, crRNA, and probe were screened in detail.…”

“…Although the sample size was inadequate to evaluate the overall validity of our method, the available data indicate that the effectiveness of this CRISPR immunochromatography method was consistent with that of the gold‐standard RT‐PCR method. 22 Notably, the CRISPR immunochromatography method can report testing results within 1 h, thus saving 30 min to 1 h when compared with RT‐PCR. For sample nucleic acid preparation, magnetic beads based quick extraction method was suitable for CRISPR detection assays, as it can provide high quality of purified nucleic acid in a relative short time.…”