Fast and Sensitive Detection of SARS-CoV-2 Nucleic Acid Using a Rapid Detection System Free of RNA Extraction

Liang Ma; Yanyan Fan; Xiaomu Kong; Yongwei Jiang; Hong Huang; Meimei Zhao; Yi Liu; Peng Gao; Yong Cui; Yongtong Cao

doi:10.1155/2023/8053524

Fast and Sensitive Detection of SARS-CoV-2 Nucleic Acid Using a Rapid Detection System Free of RNA Extraction

Int J Anal Chem. 2023 Jan 20:2023:8053524. doi: 10.1155/2023/8053524. eCollection 2023.

Authors

Liang Ma¹, Yanyan Fan², Xiaomu Kong¹, Yongwei Jiang¹, Hong Huang³, Meimei Zhao¹, Yi Liu¹, Peng Gao¹, Yong Cui⁴, Yongtong Cao¹

Affiliations

¹ Department of Clinical Laboratory, China-Japan Friendship Hospital, Beijing 100029, China.
² Department of Pulmonary & Critical Care Medicine, Laboratory of Clinical Microbiology & Infectious Diseases, China-Japan Friendship Hospital, Beijing 100029, China.
³ Coyote Bioscience USA Inc., 965, Atlantic Ave, Suite 101, Alameda, CA 94501, USA.
⁴ Department of Dermatology, China-Japan Friendship Hospital, Beijing 100029, China.

Abstract

Objectives: To establish and evaluate the analytical and clinical performance of the Flash20 SARS-CoV-2 nucleic acid rapid detection system free of RNA extraction.

Methods: The limit of detection (LoD) was determined using a negative nasopharyngeal swab matrix spiked with different concentrations of SARS-CoV-2 virus; a total of 734,337 reference sequences of viral genomes from GenBank were used for the in-silico analysis to assess the inclusivity of the assay. The specificity of the system was evaluated by testing 27 medically relevant organisms. A total of 115 clinical specimens were collected and tested on the Flash20 SARS-CoV-2 detection system and with an FDA-approved comparator test to assess the clinical performance of the system.

Results: The LoD of the Flash20 SARS-CoV-2 detection system is 250 copies/mL with a positive rate ≥90% (n = 20); alignments results showed that over 99% identity of the primer and probe of the Flash20 SARS-CoV-2 nucleic acid rapid detection system to the available SARS-CoV-2 sequences; the omicron samples tested 100% positive. None of the 27 organisms showed cross-reactivity with the Flash20 SARS-CoV-2 nucleic acid rapid detection system. Among all the 215 clinical samples, the Flash20 SARS-CoV-2 nucleic acid rapid detection system exhibits a high sensitivity of 99.24% (131/132) and 100% (83/83) specificity.

Conclusion: The nucleic acid rapid detection system provides sensitive and accurate detection of SARS-CoV-2 free of RNA extraction. The high sensitivity and short time to results of approximately 35 minutes may impact earlier infection control and disease management.