tetano
Editor, Senior Moderator
J Appl Microbiol
. 2022 Aug 21.
doi: 10.1111/jam.15788. Online ahead of print.
Flexible multiplex PCR to detect SARS-CoV-2, coronavirus OC43 and influenza A virus in nasopharyngeal swab samples
Eduardo Pelegri-Martinez[SUP] 1 [/SUP], Xabier Guruceaga[SUP] 1 [/SUP], Leire Martin-Souto[SUP] 1 [/SUP], Ana Abad-Diaz-de-Cerio[SUP] 1 [/SUP], Aitor Rementeria[SUP] 1 [/SUP], Alazne Dominguez[SUP] 2 [/SUP], Mikel Gallego[SUP] 2 [/SUP], Oscar Martinez[SUP] 2 [/SUP], Eunate Arana-Arri[SUP] 2 [/SUP], Maitane Aranzamendi[SUP] 2 [/SUP], Andoni Ramirez-Garcia[SUP] 1 [/SUP]
Affiliations
Abstract
Introduction: Quantitative Reverse Transcription PCR (RT-qPCR) is the leading tool to detect SARS-CoV-2 virus. Given that it will almost certainly continue to coexist with other respiratory viruses in the coming years, our study aimed to design a multiplex PCR system not affected by supplier outages and with reduced cost compared to the existing commercially available kits.
Methods and results: In this study, combinations of four primers/probe sets were used to construct a flexible RT-qPCR assay which is capable of discriminating between SARS-CoV-2 and the seasonal human coronavirus HCoV-OC43, or even influenza A virus. Additionally, the human RPP30 gene was used as internal control. To demonstrate the robustness of the assay, it was applied to a collection of 150 clinical samples. The results showed 100% sensitivity and specificity compared to the automatized system used at the hospital and were better when indeterminate samples were analyzed.
Conclusions: This study provides an efficient method for the simultaneous detection of SARS-CoV-2, HCoV-OC43 and influenza A virus, and its efficacy has been tested on clinical samples showing outstanding results.
Significance and impact: The multiplex RT-qPCR designed offers an accessible and economical alternative to commercial detection kits for hospitals and laboratories with limited economic resources or facing situations of supply shortage.
Keywords: Coronavirus; Influenza A virus; Multiplex RT-qPCR; OC43; SARS-CoV-2.
. 2022 Aug 21.
doi: 10.1111/jam.15788. Online ahead of print.
Flexible multiplex PCR to detect SARS-CoV-2, coronavirus OC43 and influenza A virus in nasopharyngeal swab samples
Eduardo Pelegri-Martinez[SUP] 1 [/SUP], Xabier Guruceaga[SUP] 1 [/SUP], Leire Martin-Souto[SUP] 1 [/SUP], Ana Abad-Diaz-de-Cerio[SUP] 1 [/SUP], Aitor Rementeria[SUP] 1 [/SUP], Alazne Dominguez[SUP] 2 [/SUP], Mikel Gallego[SUP] 2 [/SUP], Oscar Martinez[SUP] 2 [/SUP], Eunate Arana-Arri[SUP] 2 [/SUP], Maitane Aranzamendi[SUP] 2 [/SUP], Andoni Ramirez-Garcia[SUP] 1 [/SUP]
Affiliations
- PMID: 35988051
- DOI: 10.1111/jam.15788
Abstract
Introduction: Quantitative Reverse Transcription PCR (RT-qPCR) is the leading tool to detect SARS-CoV-2 virus. Given that it will almost certainly continue to coexist with other respiratory viruses in the coming years, our study aimed to design a multiplex PCR system not affected by supplier outages and with reduced cost compared to the existing commercially available kits.
Methods and results: In this study, combinations of four primers/probe sets were used to construct a flexible RT-qPCR assay which is capable of discriminating between SARS-CoV-2 and the seasonal human coronavirus HCoV-OC43, or even influenza A virus. Additionally, the human RPP30 gene was used as internal control. To demonstrate the robustness of the assay, it was applied to a collection of 150 clinical samples. The results showed 100% sensitivity and specificity compared to the automatized system used at the hospital and were better when indeterminate samples were analyzed.
Conclusions: This study provides an efficient method for the simultaneous detection of SARS-CoV-2, HCoV-OC43 and influenza A virus, and its efficacy has been tested on clinical samples showing outstanding results.
Significance and impact: The multiplex RT-qPCR designed offers an accessible and economical alternative to commercial detection kits for hospitals and laboratories with limited economic resources or facing situations of supply shortage.
Keywords: Coronavirus; Influenza A virus; Multiplex RT-qPCR; OC43; SARS-CoV-2.