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Comparison of Primer-Probe Sets among Different Master Mixes for Laboratory Screening of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2).
Biomed Res Int. 2020; 2020:7610678.BR

Abstract

Background

There is a shortage of chemical reagents for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnosis and a surge of SARS-CoV-2 cases, especially in limited-resource settings. Therefore, the combination of an optimal assay kit is necessary.

Methods

We compared the ability to screen SARS-CoV-2 among three primer-probe sets in two different master mixes, Invitrogen™ SuperScript™ III One-Step RT-PCR and LightCycler Multiplex RNA Virus Master.

Results

The assay with TIB-Molbiol, IDT, and Phu Sa sets for LightCycler Multiplex RNA Virus Master or Invitrogen™ SuperScript™ III One-Step RT-PCR showed positive results from a single reaction of triplicate in the three days of 4.8 copies per reaction. R squared and amplification efficiency were 0.97 and ranged from 107 to 108%, respectively.

Conclusions

Our findings indicated that TIB-Molbiol, IDT, and Phu Sa primer-probe sets could be beneficial for the laboratory screening of SARS-CoV-2 by RT-qPCR assay of E gene. There is a need to consider the combination of these reagent sets as a new strategy to increase the testing capacity of screening programs for COVID-19.

Authors+Show Affiliations

Directorial board, Pasteur Institute in Ho Chi Minh City, Vietnam.Planning Division, Pasteur Institute in Ho Chi Minh City, Vietnam.Medical Analysis Department, Pasteur Institute in Ho Chi Minh City, Vietnam.Microbiology and Immunology Department, Pasteur Institute in Ho Chi Minh City, Vietnam.Microbiology and Immunology Department, Pasteur Institute in Ho Chi Minh City, Vietnam.Microbiology and Immunology Department, Pasteur Institute in Ho Chi Minh City, Vietnam.Microbiology and Immunology Department, Pasteur Institute in Ho Chi Minh City, Vietnam.Directorial board, Pasteur Institute in Ho Chi Minh City, Vietnam.

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

33029522

Citation

Cuong, Hoang Quoc, et al. "Comparison of Primer-Probe Sets Among Different Master Mixes for Laboratory Screening of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)." BioMed Research International, vol. 2020, 2020, p. 7610678.
Cuong HQ, Hai ND, Linh HT, et al. Comparison of Primer-Probe Sets among Different Master Mixes for Laboratory Screening of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Biomed Res Int. 2020;2020:7610678.
Cuong, H. Q., Hai, N. D., Linh, H. T., Anh, N. H., Hieu, N. T., Thang, C. M., Thao, N. T. T., & Lan, P. T. (2020). Comparison of Primer-Probe Sets among Different Master Mixes for Laboratory Screening of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). BioMed Research International, 2020, 7610678. https://doi.org/10.1155/2020/7610678
Cuong HQ, et al. Comparison of Primer-Probe Sets Among Different Master Mixes for Laboratory Screening of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Biomed Res Int. 2020;2020:7610678. PubMed PMID: 33029522.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Comparison of Primer-Probe Sets among Different Master Mixes for Laboratory Screening of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). AU - Cuong,Hoang Quoc, AU - Hai,Nguyen Duc, AU - Linh,Hoang Thuy, AU - Anh,Nguyen Hoang, AU - Hieu,Nguyen Trung, AU - Thang,Cao Minh, AU - Thao,Nguyen Thi Thanh, AU - Lan,Phan Trong, Y1 - 2020/09/25/ PY - 2020/06/13/received PY - 2020/08/13/accepted PY - 2020/10/8/entrez PY - 2020/10/9/pubmed PY - 2020/10/21/medline SP - 7610678 EP - 7610678 JF - BioMed research international JO - Biomed Res Int VL - 2020 N2 - Background: There is a shortage of chemical reagents for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnosis and a surge of SARS-CoV-2 cases, especially in limited-resource settings. Therefore, the combination of an optimal assay kit is necessary. Methods: We compared the ability to screen SARS-CoV-2 among three primer-probe sets in two different master mixes, Invitrogen™ SuperScript™ III One-Step RT-PCR and LightCycler Multiplex RNA Virus Master. Results: The assay with TIB-Molbiol, IDT, and Phu Sa sets for LightCycler Multiplex RNA Virus Master or Invitrogen™ SuperScript™ III One-Step RT-PCR showed positive results from a single reaction of triplicate in the three days of 4.8 copies per reaction. R squared and amplification efficiency were 0.97 and ranged from 107 to 108%, respectively. Conclusions: Our findings indicated that TIB-Molbiol, IDT, and Phu Sa primer-probe sets could be beneficial for the laboratory screening of SARS-CoV-2 by RT-qPCR assay of E gene. There is a need to consider the combination of these reagent sets as a new strategy to increase the testing capacity of screening programs for COVID-19. SN - 2314-6141 UR - https://www.unboundmedicine.com/medline/citation/33029522/Comparison_of_Primer_Probe_Sets_among_Different_Master_Mixes_for_Laboratory_Screening_of_Severe_Acute_Respiratory_Syndrome_Coronavirus_2__SARS_CoV_2__ L2 - https://doi.org/10.1155/2020/7610678 DB - PRIME DP - Unbound Medicine ER -