Simultaneous Detection of Omicron and Other SARS-CoV-2 Variants by Multiplex PCR MassARRAY Technology.

Autor: Wacharapluesadee S; Thai Red Cross Emerging Infectious Diseases Clinical Center, King Chulalongkorn Memorial Hospital, Bangkok, Thailand., Hirunpatrawong P; Thai Red Cross Emerging Infectious Diseases Clinical Center, King Chulalongkorn Memorial Hospital, Bangkok, Thailand., Petcharat S; Thai Red Cross Emerging Infectious Diseases Clinical Center, King Chulalongkorn Memorial Hospital, Bangkok, Thailand., Torvorapanit P; Thai Red Cross Emerging Infectious Diseases Clinical Center, King Chulalongkorn Memorial Hospital, Bangkok, Thailand., Jitsatja A; Thai Red Cross Emerging Infectious Diseases Clinical Center, King Chulalongkorn Memorial Hospital, Bangkok, Thailand., Thippamom N; Thai Red Cross Emerging Infectious Diseases Clinical Center, King Chulalongkorn Memorial Hospital, Bangkok, Thailand., Ninwattana S; Thai Red Cross Emerging Infectious Diseases Clinical Center, King Chulalongkorn Memorial Hospital, Bangkok, Thailand., Phanlop C; Thai Red Cross Emerging Infectious Diseases Clinical Center, King Chulalongkorn Memorial Hospital, Bangkok, Thailand., Buathong R; Division of International Communicable Disease Control Ports and Quarantine, Department of Diseases Control, Ministry of Public Health, Nonthaburi, Thailand., Tangwangvivat R; Division of Communicable Diseases, Department of Diseases Control, Ministry of Public Health, Nonthaburi, Thailand., Klungthong C; Department of Virology, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand., Chinnawirotpisan P; Department of Virology, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand., Hunsawong T; Department of Virology, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand., Suthum K; Office of Disease Prevention and Control, Region 5, Department of Diseases Control, Ministry of Public Health, Ratchaburi, Thailand., Komolsiri S; Office of Disease Prevention and Control, Region 5, Department of Diseases Control, Ministry of Public Health, Ratchaburi, Thailand., Jones AR; Department of Virology, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand., Fernandez S; Department of Virology, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand., Putcharoen O; Thai Red Cross Emerging Infectious Diseases Clinical Center, King Chulalongkorn Memorial Hospital, Division of Infectious Diseases, Department of Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
Jazyk: angličtina
Zdroj: Research square [Res Sq] 2023 Jan 17. Date of Electronic Publication: 2023 Jan 17.
DOI: 10.21203/rs.3.rs-2482226/v1
Abstrakt: The rapid emergence of SARS-CoV-2 variants with high severity and transmutability adds further urgency for rapid and multiplex molecular testing to identify the variants. A nucleotide matrix-assisted laser-desorption-ionization time-of-flight mass spectrophotometry (MALDI-TOF MS)-based assay was developed (called point mutation array, PMA) to identify four major SARS-CoV-2 variants of concern (VOCs) including Alpha, Beta, Delta, and Omicron (namely PMA-ABDO) and differentiate Omicron subvariant (namely PMA-Omicron). PMA-ABDO and PMA-Omicron consist of 24 and 28 mutation sites of the spike gene. Both PMA panels specifically identified VOCs with as low as 10 viral copies/ µl. The panel has shown a 100% concordant with the Next Generation Sequencing (NGS) results testing on 256 clinical specimens with real-time PCR cycle threshold (Ct) values less than 26. It showed a higher sensitivity over NGS; 25/28 samples were positive by PMA but not NGS in the clinical samples with PCR Ct higher than 26. Due to the mass of nucleotide used to differentiate between wild-type and mutation strains, the co-infection or recombination of multiple variants can be determined by the PMA method. This method is flexible in adding a new primer set to identify a new emerging mutation site among the current circulating VOCs and the turnaround time is less than 8 hours. However, the spike gene sequencing or NGS retains the advantage of detecting newly emerged variants.
Databáze: MEDLINE
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