ArticleJournal of clinical microbiology2005
Automated extraction of viral-pathogen RNA and DNA for high-throughput quantitative real-time PCR.
Article in Journal of clinical microbiology, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed.
- Colorimetric determination of Listeria monocytogenes using aptamer and urease dual-labeled magnetic nanoparticles and cucurbit[7]uril-mediated supramolecular assembly of gold nanoparticle.Mikrochimica acta · 2021Article
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- Epstein-Barr virus in peripheral blood is associated with response to rituximab therapy in rheumatoid arthritis patients.Clinical rheumatology · 2015Article
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- Development of a low-resource RNA extraction cassette based on surface tension valves.ACS applied materials & interfaces · 2011Article
- Performance characteristics of the MultiCode-RTx hepatitis C virus load test targeting the 3' untranslated region.Journal of clinical microbiology · 2010Article
- Comparisons of three automated systems for genomic DNA extraction in a clinical diagnostic laboratory.Yonsei medical journal · 2010Article
- Comparative evaluation of a commercially available automated system for extraction of viral DNA from whole blood: application to monitoring of epstein-barr virus and cytomegalovirus load.Journal of clinical microbiology · 2009Article
- Clinical predictive value of real-time PCR quantification of human cytomegalovirus DNA in amniotic fluid samples.Journal of clinical microbiology · 2009Article
- Detection of perinatal cytomegalovirus infection and sensorineural hearing loss in belgian infants by measurement of automated auditory brainstem response.Journal of clinical microbiology · 2008Article
- Comparison of automated nucleic acid extraction methods with manual extraction.The Journal of molecular diagnostics : JMD · 2008Article
- Performance characteristics of a quantitative hepatitis C virus RNA assay using COBAS AmpliPrep total nucleic acid isolation and COBAS taqman hepatitis C virus analyte-specific reagent.The Journal of molecular diagnostics : JMD · 2008Article
- Evaluation of the Epstein-Barr virus R-gene quantification kit in whole blood with different extraction methods and PCR platforms.The Journal of molecular diagnostics : JMD · 2008Article
- Evaluation of real-time PCR laboratory-developed tests using analyte-specific reagents for cytomegalovirus quantification.Journal of clinical microbiology · 2007Article
- Evaluation of NucliSens easyMAG for automated nucleic acid extraction from various clinical specimens.Journal of clinical microbiology · 2007Article
- Expertise of laboratories in viral load quantification, genotyping, and precore mutant determination for hepatitis B virus in a multicenter study.Journal of clinical microbiology · 2006Article
- Fluctuating hepatitis B viremia: Full of sound and fury, signifying nothing?Saudi journal of gastroenterology : official journal of the Saudi Gastroenterology AssociationArticle
Corrections and comments
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The performance of the m1000 system (Abbott Laboratories, Illinois) as a front-end extraction system for high-throughput "in-house" quantitative real-time PCR assays was analyzed and compared to that of manual extraction of plasma and serum samples (hepatitis C virus [HCV] and hepatitis B virus [HBV]) and EDTA-blood samples (cytomegalovirus [CMV] and Epstein-Barr virus [EBV]). Linearity of extraction was tested on dilution series of HCV and HBV reference materials. The correlation coefficient for standard curves based on repeated extraction runs was 0.97 +/- 0.06 for HCV and 0.97 +/- 0.03 for HBV, indicating a linear extraction from 100 to 1.0 x 10(5) HCV IU/ml and from 100 to 1.0 x 10(6) HBV IU/ml. Intra- and interrun variability was below 0.23 log(10) IU/ml for 2.98 to 5.28 log(10) HCV IU/ml and 2.70 to 5.20 log(10) HBV IU/ml. Correlation between automated and manual extraction was very good. For HCV, the correlation coefficient was 0.91 and the mean difference in viral load was 0.13 log(10) HCV IU/ml. For HBV, the correlation coefficient was 0.98 and the mean difference in viral load 0.61 log(10) HBV IU/ml. For CMV and EBV, the correlation coefficient was 0.98 and the mean difference in viral load 0.33 log(10) copies/ml. Accuracy was confirmed with a reference panel (QCMD, Glasgow, Scotland) for all four assays. No cross-contamination was observed when extracting strongly positive polyomavirus samples (8.10 log(10) copies/ml) interspersed with polyomavirus-negative samples. Automated extraction via the m1000 system offers a high reliability of extraction and resulted in a strong reduction of the required extraction hands-on time for high-throughput PCR compared to manual extraction protocols.
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