Comparison of five commercial DNA extraction kits for the recovery of Yersinia pestis DNA from bacterial suspensions and spiked environmental samples

Authors

  • L.A. Dauphin,

    1.  Bioterrorism Rapid Response and Advanced Technology (BRRAT) Laboratory, Division of Bioterrorism Preparedness and Response (DBPR), National Center for Preparedness, Detection, and Control of Infectious Diseases (NCPDCID), Centers for Disease Control and Prevention (CDC), Atlanta, GA, USA
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  • K.W. Stephens,

    1.  Bioterrorism Rapid Response and Advanced Technology (BRRAT) Laboratory, Division of Bioterrorism Preparedness and Response (DBPR), National Center for Preparedness, Detection, and Control of Infectious Diseases (NCPDCID), Centers for Disease Control and Prevention (CDC), Atlanta, GA, USA
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  • S.C. Eufinger,

    1.  Bioterrorism Rapid Response and Advanced Technology (BRRAT) Laboratory, Division of Bioterrorism Preparedness and Response (DBPR), National Center for Preparedness, Detection, and Control of Infectious Diseases (NCPDCID), Centers for Disease Control and Prevention (CDC), Atlanta, GA, USA
    2.  Graduate Division of Biological and Biomedical Sciences, Emory University, Atlanta, GA, USA
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  • M.D. Bowen

    1.  Bioterrorism Rapid Response and Advanced Technology (BRRAT) Laboratory, Division of Bioterrorism Preparedness and Response (DBPR), National Center for Preparedness, Detection, and Control of Infectious Diseases (NCPDCID), Centers for Disease Control and Prevention (CDC), Atlanta, GA, USA
    2.  Gastroenteritis and Respiratory Viruses Laboratory Branch, Division of Viral Diseases, National Center for Immunization and Respiratory Diseases, CDC, Atlanta, GA, USA
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Leslie Ann Dauphin, BRRAT Laboratory, DBPR, NCPDCID, CDC, Mail Stop G-42, 1600 Clifton Road, Atlanta, GA 30333, USA. E-mail: Ldauphin@CDC.GOV

Abstract

Aim:  To evaluate commercial DNA extraction kits for their ability to isolate DNA from Yersinia pestis suspensions and spiked environmental samples.

Methods and Results:  Five commercially available DNA extraction kits were evaluated: the ChargeSwitch gDNA Mini Bacteria Kit, the IT 1-2-3 Sample DNA Purification Kit, the MasterPure Complete DNA and RNA Purification Kit, the QIAamp DNA Blood Mini Kit and the UltraClean Microbial DNA Isolation Kit. The extraction methods were performed upon six Y. pestis strains and spiked environmental specimens, including three swab types and one powder type. Taqman real-time PCR analysis revealed that the use of the MasterPure kit resulted in DNA with the most consistently positive results and the lowest limit of detection from Y. pestis suspensions and spiked environmental samples.

Conclusion:  Comparative evaluations of the five commercial DNA extraction methods indicated that the MasterPure kit was superior for the isolation of PCR-amplifiable DNA from Y. pestis suspensions and spiked environmental samples.

Significance and Impact of the Study:  The results of this study can assist diagnostic laboratories with selecting the best extraction method for processing environmental specimens for subsequent detection of Y. pestis by real-time PCR.

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