Literature DB >> 22750394

Comparison of nucleic acid extraction platforms for detection of select biothreat agents for use in clinical resource limited settings.

Michelle A Shipley1, Jeffrey W Koehler, David A Kulesh, Timothy D Minogue.   

Abstract

High-quality nucleic acids are critical for optimal PCR-based diagnostics and pathogen detection. Rapid sample processing time is important for the earliest administration of therapeutic and containment measures, especially in the case of biothreat agents. In this context, we compared the Fujifilm QuickGene-Mini80 to Qiagen's QIAamp Mini Purification kits for extraction of DNA and RNA for potential use in austere settings. Qiagen (QIAamp) column-based extraction is the currently recommended purification platform by United States Army Medical Research Institute for Infectious Diseases for both DNA and RNA extraction. However, this sample processing system requires dedicated laboratory equipment including a centrifuge. In this study, we investigated the QuickGene-Mini80, which does not require centrifugation, as a suitable platform for nucleic acid extraction for use in resource-limited locations. Quality of the sample extraction was evaluated using pathogen-specific, real-time PCR assays for nucleic acids extracted from viable and γ-irradiated Bacillus anthracis, Yersinia pestis, vaccinia virus, Venezuelan equine encephalitis virus, or B. anthracis spores in buffer or human whole blood. QuickGene-Mini80 and QIAamp performed similarly for DNA extraction regardless of organism viability. It was noteworthy that γ-irradiation did not have a significant impact on real-time PCR for organism detection. Comparison with QIAamp showed a less than adequate performance of the Fujifilm instrument for RNA extraction. However, QuickGene-Mini80 remains a viable alternative to QIAamp for DNA extraction for use in remote settings due to extraction quality, time efficiency, reduced instrument requirements, and ease of use. Published by Elsevier B.V.

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Year:  2012        PMID: 22750394     DOI: 10.1016/j.mimet.2012.06.008

Source DB:  PubMed          Journal:  J Microbiol Methods        ISSN: 0167-7012            Impact factor:   2.363


  8 in total

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Authors:  Christopher P Stefan; Jeffrey W Koehler; Timothy D Minogue
Journal:  Sci Rep       Date:  2016-05-13       Impact factor: 4.379

5.  Development of a recombinase polymerase amplification assay for rapid detection of Francisella noatunensis subsp. orientalis.

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Journal:  PLoS One       Date:  2018-02-14       Impact factor: 3.240

6.  Evaluation of inhibitor-resistant real-time PCR methods for diagnostics in clinical and environmental samples.

Authors:  Adrienne Trombley Hall; Ashley McKay Zovanyi; Deanna Rose Christensen; Jeffrey William Koehler; Timothy Devins Minogue
Journal:  PLoS One       Date:  2013-09-09       Impact factor: 3.240

7.  An integrated lab-on-chip for rapid identification and simultaneous differentiation of tropical pathogens.

Authors:  Jeslin J L Tan; Monica Capozzoli; Mitsuharu Sato; Wanitda Watthanaworawit; Clare L Ling; Marjorie Mauduit; Benoît Malleret; Anne-Charlotte Grüner; Rosemary Tan; François H Nosten; Georges Snounou; Laurent Rénia; Lisa F P Ng
Journal:  PLoS Negl Trop Dis       Date:  2014-07-31

8.  Persistence of Zika virus in conjunctival fluid of convalescence patients.

Authors:  Jeslin J L Tan; Praveen K Balne; Yee-Sin Leo; Louis Tong; Lisa F P Ng; Rupesh Agrawal
Journal:  Sci Rep       Date:  2017-09-11       Impact factor: 4.379

  8 in total

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