About: Automated extraction protocol for quantification of SARS-Coronavirus RNA in serum: an evaluation study

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About: Automated extraction protocol for quantification of SARS-Coronavirus RNA in serum: an evaluation study 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
title	Automated extraction protocol for quantification of SARS-Coronavirus RNA in serum: an evaluation study
Creator	Jin, Yongjie Lim, Wilina Chan, Anthony Chiu, Rossa Chung, Grace Lo, Ym Lui, Wing-Bong
source	Medline; PMC
abstract	BACKGROUND: We have previously developed a test for the diagnosis and prognostic assessment of the severe acute respiratory syndrome (SARS) based on the detection of the SARS-coronavirus RNA in serum by real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR). In this study, we evaluated the feasibility of automating the serum RNA extraction procedure in order to increase the throughput of the assay. METHODS: An automated nucleic acid extraction platform using the MagNA Pure LC instrument (Roche Diagnostics) was evaluated. We developed a modified protocol in compliance with the recommended biosafety guidelines from the World Health Organization based on the use of the MagNA Pure total nucleic acid large volume isolation kit for the extraction of SARS-coronavirus RNA. The modified protocol was compared with a column-based extraction kit (QIAamp viral RNA mini kit, Qiagen) for quantitative performance, analytical sensitivity and precision. RESULTS: The newly developed automated protocol was shown to be free from carry-over contamination and have comparable performance with other standard protocols and kits designed for the MagNA Pure LC instrument. However, the automated method was found to be less sensitive, less precise and led to consistently lower serum SARS-coronavirus concentrations when compared with the column-based extraction method. CONCLUSION: As the diagnostic efficiency and prognostic value of the serum SARS-CoV RNA RT-PCR test is critically associated with the analytical sensitivity and quantitative performance contributed both by the RNA extraction and RT-PCR components of the test, we recommend the use of the column-based manual RNA extraction method.
has issue date	2006-02-09(xsd:dateTime)
bibo:doi	10.1186/1471-2334-6-20
bibo:pmid	16466582
has license	cc-by
sha1sum (hex)	882b4b4a41b772d31e8f9e860882088336f26e26
schema:url	https://doi.org/10.1186/1471-2334-6-20
resource representing a document's title	Automated extraction protocol for quantification of SARS-Coronavirus RNA in serum: an evaluation study
has PubMed Central identifier	PMC1382231
has PubMed identifier	16466582
schema:publication	BMC Infect Dis
resource representing a document's body	covid:882b4b4a41b772d31e8f9e860882088336f26e26#body_text
is schema:about of	named entity 'polymerase' named entity 'QUANTIFICATION' named entity 'INFECTIOUS DISEASES' covid:arg/882b4b4a41b772d31e8f9e860882088336f26e26 named entity 'BASED' named entity 'SARS' named entity 'ASSESSMENT' named entity 'PCR' named entity 'THROUGHPUT' named entity 'FEASIBILITY' named entity 'RNA EXTRACTION PROCEDURE' named entity 'ASSAY' named entity 'EVALUATION STUDY' named entity 'HAVE' named entity 'SERUM' named entity 'EXTRACTION PROTOCOL' named entity 'AUTOMATED' named entity 'AUTOMATING' named entity 'SERUM' named entity 'EVALUATED' named entity 'QUANTITATIVE REVERSE TRANSCRIPTASE POLYMERASE CHAIN REACTION' named entity 'DIAGNOSIS' named entity 'REAL-TIME' named entity 'TO INCREASE' named entity 'TEST' named entity 'STUDY' named entity 'SARS' named entity 'BMC' named entity 'SEVERE ACUTE RESPIRATORY SYNDROME' named entity 'PROGNOSTIC' named entity 'ORDER' named entity 'PREVIOUSLY' named entity 'DETECTION' named entity 'CORONAVIRUS RNA' named entity 'chain reaction' named entity 'real-time' named entity 'throughput' named entity 'BMC Infectious Diseases' named entity 'RT-PCR' named entity 'prognostic assessment' named entity 'serum' named entity 'coronavirus' named entity 'SARS' named entity 'severe acute respiratory syndrome' named entity 'BMC' named entity 'SARS' named entity 'RNA' named entity 'lysis' named entity 'serially diluted' named entity 'silica' named entity 'RT-PCR' named entity 'Transport medium' named entity 'Sequence Detector' named entity 'SARS-CoV' named entity 'Roche Diagnostics' named entity 'Viral' named entity 'SARS-CoV' named entity 'biosafety' named entity 'RNA' named entity 'prognostic' named entity 'SARS-CoV' named entity 'lysis' named entity 'regression line' named entity 'severe acute respiratory syndrome' named entity 'proteinase' named entity 'RNA' named entity 'reverse transcriptase polymerase chain reaction' named entity 'SARS-CoV' named entity 'SARS-CoV' named entity 'SARS-CoV' named entity 'pathogens' named entity 'plasma' named entity 'lysis buffer' named entity 'epidemic' named entity 'Foster City'

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