A NOVEL ULTRAFAST REAL-TIME PCR INSTRUMENT FOR RAPID DETECTION OF SARS-COV-2 NUCLEIC ACIDS IN 30 MINUTES

ABSTRACT

Intro The coronavirus disease 2019 (COVID-19) pandemic is still raging around the world. Therefore, an efficient, rapid, and low-cost nucleic acid detection method for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is urgently needed. There is a miniaturized ultra-fast real-time PCR instrument, and we will verify and evaluate its 30-minute rapid detection of SARS-CoV-2. Methods A rapid PCR temperature change mode was explored by moving the reaction tube between the independent temperature modules with large temperature differences and a portable ultra-fast real-time PCR instrument were developed. We established a rapid SARS-CoV-2 test method using the instrument, a China Food and Drug Administration-certified SARS-CoV-2 reagent and optimized reaction condition. The analytical and clinical performances of the rapid tests were evaluated by comparing with the standard SARS-CoV-2 tests. Findings The new temperature change mode can effectively shorten the amplification reaction time. The rapid SARS-CoV-2 test method was established and the time to yield results were greatly shortened from 81 min of the standard test to 31 min. Specificity of the rapid test was assessed and no non-specific amplification (0/63) was observed. Clinical performance was evaluated using 184 respiratory specimens from patients with suspected SARS-CoV-2 infection. The positive agreement between the rapid and standard tests was 100% (67/67), the negative agreement was 97.4% (114/117), and the kappa statistic was 0.965 (P<0.001). No significant differences in the Ct values for each target gene were observed between the rapid test and the standard test (P>0.05). Conclusion A 30-minute detection method for SARS-CoV-2 nucleic acid using a novel ultra-fast real-time PCR instrument was developed. The waiting time for test results in cases of suspected infection may be reduced greatly by using the ultra-fast real-time PCR instrument