Optimal stabilization for long-term storage of nucleic acid-based CRISPR/Cas12a assay for SARS-CoV-2 detection

Chonticha Saisawang, Molecular Medical Biosciences Cluster, Institute of Molecular Biosciences, Mahidol University, Salaya Campus, Salaya 73170, ThailandFollow
Piyaporn Naksith, Molecular Medical Biosciences Cluster, Institute of Molecular Biosciences, Mahidol University, Salaya Campus, Salaya 73170, Thailand
Somsri Sakdee, Molecular Medical Biosciences Cluster, Institute of Molecular Biosciences, Mahidol University, Salaya Campus, Salaya 73170, Thailand
Albert J. Ketterman, Molecular Medical Biosciences Cluster, Institute of Molecular Biosciences, Mahidol University, Salaya Campus, Salaya 73170, Thailand
Soontorn Tuntithavornwat, Department of Chemical Engineering, Faculty of Engineering, Mahidol University, Salaya Campus, Salaya 73170, Thailand
Pattaraporn Nimsamer, Research Unit of Systems Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
Oraphan Mayuramart, Research Unit of Systems Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
Naphat Chantaravisoot, Center of Excellence in Systems Biology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
Trairak Pisitkun, Center of Excellence in Systems Biology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
Sunchai Payungporn, Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand

Abstract

In this long-term storage study, we optimized the lyophilization conditions of each reaction stage of a nucleic acid-based assay for SARS-CoV-2 detection. The stability testing demonstrated that the dried reactions from all 3 steps (cDNA synthesis, isothermal amplification and detection) can be kept at -20°C or 4°C for up to 6 or 3 months, respectively, whereas, if stored at 25°C or 37°C, the reagents only could be stored for a few days without quality loss. This suggests that we can have the dried reactions at -20°C for long-term storage until needed. Moreover, this assay is now simpler to perform as each of the 3 steps now proceeds with pre-mixed regents lyophilized in a single tube for each step.

Recommended Citation

Saisawang, Chonticha; Naksith, Piyaporn; Sakdee, Somsri; Ketterman, Albert J.; Tuntithavornwat, Soontorn; Nimsamer, Pattaraporn; Mayuramart, Oraphan; Chantaravisoot, Naphat; Pisitkun, Trairak; and Payungporn, Sunchai (2023) "Optimal stabilization for long-term storage of nucleic acid-based CRISPR/Cas12a assay for SARS-CoV-2 detection," Karbala International Journal of Modern Science: Vol. 9 : Iss. 2 , Article 4.
Available at: https://doi.org/10.33640/2405-609X.3288