Other than clinical samples, what can be used as a positive control to validate custom qPCR or PCR assay for COVID19?
For preliminary screening, what can be used as positive control to validate custom qPCR or PCR assay? any spike protein cDNA expression plasmid available?
I understand clinical samples would be the final confirmation.
You can order COVID-19 RNA that can work as your positive control. Here a list of sites where you can order the sequences, just make sure they match the area of your primers and probe:
https://ec.europa.eu/jrc/en/news/new-control-material-developed-jrc-scientists-help-prevent-coronavirus-test-failures
https://crm.jrc.ec.europa.eu/p/EURM-019
https://www.european-virus-archive.com/
https://www.european-virus-archive.com/nucleic-acid/sars-cov-frankfurt-1-rna
https://www.european-virus-archive.com/detection-kit/2019-ncov-e-gene-stabilized-rna-positive-control-shipping-room-temperature
Please refer to this article, where the authors ordered positive controls online. Thanks
Article First confirmed detection of SARS-CoV-2 in untreated wastewa...
Real-time qPCR for COVID-19.
you can refer to this.
https://www.takarabio.com/products/covid-19-research/viral-detection-with-qpcr/viral-detection-via-qpcr
You can order some target oligo sequences to use as positive control. This company is making some RNA sequences for us to use as positive control. As the products are RNA, these are safe to use in any laboratory: https://eu.idtdna.com/pages/products/custom-dna-rna/custom-rna-oligos
NIBSC, European virus archive global and many others provide SARS CoV2 controls.
https://www.european-virus-archive.com/
For preliminary screening using qPCR, you shall have a positive control that chemically synthesis for your target genes (e.g. E Sarbeco gene or RdRP gene) to validate the qPCR reagents that used to run your screening samples in the same run. On top of that, it will be even better to have internal control that target human gene (e.g. RNAse P gene) as what CDC assay did, which typically to make sure the swap collection was done correctly in getting sufficient total RNA of human for the qPCR screening.
Our company is assisting both academic and medical researchers in establishing their qPCR screening for COVID-19. Before this, we used to offer customized qPCR screening/ expression detection services under our Molecular Biology Services. From time to time, researchers are looking for our services in their qPCR assay optimization and resolving the problems associated with their screening assays.
We thus developed our very first version of COVID-19 positive control for qPCR screening assay to include only E_Sarbeco gene, and RdRP gene according to Charité/Berlin qPCR protocol (version 17th Jan 2020). Along the time, we found researchers need more than these two genes and some of them struggled with the NTC problems from this assay.
We thus developed second version of this COVID-19 positive control for qPCR screening assay to add new fragments: (i) RdRP Helicase, and (ii) human RNAse P; according to Chan JF-W et al., 2020 and CDC assays for SARS-CoV-2 detection respectively. As mentioned, Human RNAse P gene is used as internal control (IC) for qPCR assay of SARS-CoV-2 detection. Furthermore, we have introduced a pair of modified qPCR primers (Product No: CV013 and CV014) to better detect RdRP gene using the same RdRP P2 probe in Charité/Berlin protocol. This resolves the assay sensitivity to better distinguish the signal between NTC and RdRP gene.
For more information, you may reach us at https://base-asia.com/covid-19-solutions