With the shortage of reagents for molecular tests worldwide we are looking for a protocol that doesn`t need to have an extraction step and that can directly detect the SARS-CoV-2 virus using a RT-Real-Time PCR protocol for respiratory samples. We are very thankful for any information, reference or protocols shared.
These might be of interest to you:
1. Article Digital PCR for direct quantification of viruses without DNA...
2. https://academic.oup.com/biomethods/article/2/1/bpx008/3858804
The first is for viral DNA detection; the second is for viral RNA and is reported to not require standard reverse transcription or RNA extraction
Thank you for your work
J. Mandula
Thank you kindly Jessica. I will look carefully the links you sent. Best regards
Marcelo
Ola Marcelo, tudo bem? Sou Prof da UFPR em Matinhos. Tenho bastante experiência na área de biologia molecular, proteínas etc... Me coloco a disposição, junto com a minha equipe, para ajudar no que for preciso como por exemplo procurando metodologias alternativas, procurando informações bibliográficas, etc....Também dispomos de um pequeno lab de microbiologia biologia molecular em Matinhos apto a realizar microbiologia molecular básica PCR, eletroforese etc,,,infelizmente não temos qRT-PCR. De qualquer forma nossa estrutura está a sua disposição. Também tenho contato com colegas em Curitiba com laboratórios mais equipados que podem ser úteis e eventualmente tenham interesse em ajudar
whats: +55 41 996765856
abraços
Dear Marcello, I have the same question here in Brasília. We have the PCR kits and Kits for manual RNA extraction, but we really prefer to reduce manipulations of clinical samples with SARS-CoV-2. See the following articles:
1-Preliminary support for a “dry swab, extraction free” protocol for SARS-CoV-2 testing via RT-qPCR. Sanjay Srivatsan. bioRxiv preprint doi: https://doi.org/10.1101/2020.04.22.056283.
2-SARS-CoV-2 detection from nasopharyngeal swab samples without RNA extraction. Carolina Beltrán-Pavez et al. bioRxiv preprint doi: https://doi.org/10.1101/2020.03.28.013508.
3-Detection of SARS-CoV-2 RNA by direct RT-qPCR on nasopharyngeal specimens without extraction of viral RNA. Mohammad Rubayet Hasan et al. medRxiv preprint doi: https://doi.org/10.1101/2020.04.18.20070755.
4- An alternative workflow for molecular detection of SARS-CoV-2 - escape from the NA extraction kit-shortage, Copenhagen, Denmark, March 2020. Anna S. Fomsgaard et al. . doi: https://doi.org/10.1101/2020.03.27.20044495 medRxiv preprint
5-Optimization of SARS-CoV-2 detection by RT-QPCR without RNA extraction
April 2020 DOI: 10.1101/2020.04.06.028902. Natacha Merindol et al.
I'll try some protocols this week!
All the best,
Ricardo Titze
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