About: SYBR green one-step qRT-PCR for the detection of SARS-CoV-2 RNA in saliva

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About: SYBR green one-step qRT-PCR for the detection of SARS-CoV-2 RNA in saliva Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : covidontheweb.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	SYBR green one-step qRT-PCR for the detection of SARS-CoV-2 RNA in saliva
Creator	Aa, Millar Ab, Smith Am, Fahrer Dr, Ganguly Ganguly, Diep Nc, Khin Wy, Hee
Source	BioRxiv
abstract	We describe our efforts at developing a one-step quantitative reverse-transcription (qRT)-PCR protocol to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA directly from saliva samples, without RNA purification. We find that both heat and the presence of saliva impairs the ability to detect synthetic SARS-CoV-2 RNA. Buffer composition (for saliva dilution) was also crucial to effective PCR detection. Using the SG2 primer pair, designed by Sigma-Aldrich, we were able to detect the equivalent of 1.7×106 viral copies per mL of saliva after heat inactivation; approximately equivalent to the median viral load in symptomatic patients. This would make our assay potentially useful for rapid detection of high-shedding infected individuals. We also provide a comparison of the PCR efficiency and specificity, which varied considerably, across 9 reported primer pairs for SARS-CoV-2 detection. Primer pairs SG2 and CCDC-N showed highest specificity and PCR efficiency. Finally, we provide an alternate primer pair to use as a positive control for human RNA detection in SARS-CoV-2 assays, as we found that the widely used US CDC primers (targeting human RPP30) do not span an exon-exon junction and therefore does not provide an adequate control for the reverse transcription reaction.
has issue date	2020-06-19(xsd:dateTime)
bibo:doi	10.1101/2020.05.29.109702
has license	biorxiv
sha1sum (hex)	83051cc3f6e7b0ee0937ca9147873f38669944d9
schema:url	https://doi.org/10.1101/2020.05.29.109702
resource representing a document's title	SYBR green one-step qRT-PCR for the detection of SARS-CoV-2 RNA in saliva
schema:publication	bioRxiv
resource representing a document's body	covid:83051cc3f6e7b0ee0937ca9147873f38669944d9#body_text
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