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[Comparative assessment of DNA extraction methods for identification of glanders and melioidosis etiological agents by PCR].

Abstract

Pathogenic Burkholderia are considered as a cause of dangerous infections and potential agents of bioterrorism. Comparative assessment of different methods of extraction and purification of DNA for PCR analysis of pure cultures and samples contaminated by etiological agents of glanders and melioidosis was performed. Samples of soil and food artificially contaminated by pathogenic Burkholderia as well as organs of infected animals were tested. DNA was extracted by methods of boiling, nucleosorption with presence of guanidine thiocyanate, guanidine thiocyanatephenol extraction, guanidine thiocyanate-phenol extraction with additional purification of DNA by nucleosorption. Amplification was performed by "Flash" technique and detector of fluorescence was used for analysis of PCR products. Utilization of the recommended methods of preparation depending on the nature of sample let to detect by the "Flash" technique the etiological agents of glanders and melioidosis in concentration =10(3) microbial cells per ml. Choice of DNA extraction and purification methods is determined by type of a sample and presence in it of admixtures inhibiting PCR.

Authors

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Pub Type(s)

Comparative Study
English Abstract
Journal Article

Language

rus

PubMed ID

18368754

Citation

Zinchenko, O V., et al. "[Comparative Assessment of DNA Extraction Methods for Identification of Glanders and Melioidosis Etiological Agents By PCR]." Zhurnal Mikrobiologii, Epidemiologii I Immunobiologii, 2008, pp. 55-60.
Zinchenko OV, Antonov VA, Tkachenko GA, et al. [Comparative assessment of DNA extraction methods for identification of glanders and melioidosis etiological agents by PCR]. Zh Mikrobiol Epidemiol Immunobiol. 2008.
Zinchenko, O. V., Antonov, V. A., Tkachenko, G. A., Altukhova, V. V., Zamaraev, V. S., Piven', N. N., Goloseev, I. u. A., Vasil'ev, V. P., Lomova, L. V., & Alekseev, V. V. (2008). [Comparative assessment of DNA extraction methods for identification of glanders and melioidosis etiological agents by PCR]. Zhurnal Mikrobiologii, Epidemiologii I Immunobiologii, (1), 55-60.
Zinchenko OV, et al. [Comparative Assessment of DNA Extraction Methods for Identification of Glanders and Melioidosis Etiological Agents By PCR]. Zh Mikrobiol Epidemiol Immunobiol. 2008 Jan-Feb;(1)55-60. PubMed PMID: 18368754.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Comparative assessment of DNA extraction methods for identification of glanders and melioidosis etiological agents by PCR]. AU - Zinchenko,O V, AU - Antonov,V A, AU - Tkachenko,G A, AU - Altukhova,V V, AU - Zamaraev,V S, AU - Piven',N N, AU - Goloseev,Iu A, AU - Vasil'ev,V P, AU - Lomova,L V, AU - Alekseev,V V, PY - 2008/3/29/pubmed PY - 2008/5/10/medline PY - 2008/3/29/entrez SP - 55 EP - 60 JF - Zhurnal mikrobiologii, epidemiologii i immunobiologii JO - Zh Mikrobiol Epidemiol Immunobiol IS - 1 N2 - Pathogenic Burkholderia are considered as a cause of dangerous infections and potential agents of bioterrorism. Comparative assessment of different methods of extraction and purification of DNA for PCR analysis of pure cultures and samples contaminated by etiological agents of glanders and melioidosis was performed. Samples of soil and food artificially contaminated by pathogenic Burkholderia as well as organs of infected animals were tested. DNA was extracted by methods of boiling, nucleosorption with presence of guanidine thiocyanate, guanidine thiocyanatephenol extraction, guanidine thiocyanate-phenol extraction with additional purification of DNA by nucleosorption. Amplification was performed by "Flash" technique and detector of fluorescence was used for analysis of PCR products. Utilization of the recommended methods of preparation depending on the nature of sample let to detect by the "Flash" technique the etiological agents of glanders and melioidosis in concentration =10(3) microbial cells per ml. Choice of DNA extraction and purification methods is determined by type of a sample and presence in it of admixtures inhibiting PCR. SN - 0372-9311 UR - https://www.unboundmedicine.com/medline/citation/18368754/[Comparative_assessment_of_DNA_extraction_methods_for_identification_of_glanders_and_melioidosis_etiological_agents_by_PCR]_ DB - PRIME DP - Unbound Medicine ER -