ArticleJournal of clinical microbiology2009
Multicenter comparative evaluation of five commercial methods for toxoplasma DNA extraction from amniotic fluid.
Article in Journal of clinical microbiology, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed, 46 citations in OpenAlex.
- Performance evaluation of nine reference centers and comparison of DNA extraction protocols for effective surveillance of Leishmania-infected Phlebotomine sand flies: Basis for technical recommendations.PLoS neglected tropical diseases · 2024Article
- Evaluation of DNA Extraction Methods for Reliable Quantification ofBiosensors · 2023Article
- Investigation of the effects of mir-219-1 gene variants on the development of disease in non-small cell lung cancer patients.African health sciences · 2022Article
- An evolving approach to the laboratory assessment of COVID-19.Journal of medical virology · 2020Review
- Multicenter Comparative Study of SixMicroorganisms · 2020Article
- Evaluation of Toxoplasma ELITe MGB Real-Time PCR Assay for Diagnosis of Toxoplasmosis.Journal of clinical microbiology · 2017Article
- Molecular diagnosis of toxoplasmosis in immunocompromised patients: a 3-year multicenter retrospective study.Journal of clinical microbiology · 2015Article
- Comparison of an automated nucleic acid extraction system with the column-based procedure.European journal of microbiology & immunology · 2015Article
- Multicentric comparative assessment of the bio-evolution Toxoplasma gondii detection kit with eight laboratory-developed PCR assays for molecular diagnosis of congenital toxoplasmosis.Journal of clinical microbiology · 2015Article
- Characterization and multicentric validation of a common standard for Toxoplasma gondii detection using nucleic acid amplification assays.Journal of clinical microbiology · 2014Article
- Comparative assessment of automated nucleic acid sample extraction equipment for biothreat agents.Journal of clinical microbiology · 2014Article
- Toxoplasma seroconversion with negative or transient immunoglobulin M in pregnant women: myth or reality? A French multicenter retrospective study.Journal of clinical microbiology · 2013Article
- The Toxoplasma MAG1 peptides induce sex-based humoral immune response in mice and distinguish active from chronic human infection.Microbes and infection · 2013Article
- Optimization of Toxoplasma gondii DNA extraction from amniotic fluid using NucliSENS easyMAG and comparison with QIAamp DNA minikit.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2012Article
- Epidemiology of and diagnostic strategies for toxoplasmosis.Clinical microbiology reviews · 2012Review
- Multicentric comparative analytical performance study for molecular detection of low amounts of Toxoplasma gondii from simulated specimens.Journal of clinical microbiology · 2010Article
Corrections and comments
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Authors and funding
6 authors at 5 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Over the past few years, a number of new nucleic acid extraction methods and extraction platforms using chemistry combined with magnetic or silica particles have been developed, in combination with instruments to facilitate the extraction procedure. The objective of the present study was to investigate the suitability of these automated methods for the isolation of Toxoplasma gondii DNA from amniotic fluid (AF). Therefore, three automated procedures were compared to two commercialized manual extraction methods. The MagNA Pure Compact (Roche), BioRobot EZ1 (Qiagen), and easyMAG (bioMérieux) automated procedures were compared to two manual DNA extraction kits, the QIAamp DNA minikit (Qiagen) and the High Pure PCR template preparation kit (Roche). Evaluation was carried out with two specific Toxoplasma PCRs (targeting the 529-bp repeat element), inhibitor search PCRs, and human beta-globin PCRs. The samples each consisted of 4 ml of AF with or without a calibrated Toxoplasma gondii RH strain suspension (0, 1, 2.5, 5, and 25 tachyzoites/ml). All PCR assays were laboratory-developed real-time PCR assays, using either TaqMan or fluorescent resonance energy transfer probes. A total of 1,178 PCRs were performed, including 978 Toxoplasma PCRs. The automated and manual methods were similar in sensitivity for DNA extraction from T. gondii at the highest concentration (25 Toxoplasma gondii cells/ml). However, our results showed that the DNA extraction procedures led to variable efficacy in isolating low concentrations of tachyzoites in AF samples (<5 Toxoplasma gondii cells/ml), a difference that might have repercussions since low parasite concentrations in AF exist and can lead to congenital toxoplasmosis.
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