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Multiplex qPCR for reliable detection and differentiation of Burkholderia mallei and Burkholderia pseudomallei.
BMC Infect Dis. 2013 Feb 14; 13:86.BI

Abstract

BACKGROUND

Burkholderia mallei and B. pseudomallei are two closely related species of highly virulent bacteria that can be difficult to detect. Pathogenic Burkholderia are endemic in many regions worldwide and cases of infection, sometimes brought by travelers from unsuspected regions, also occur elsewhere. Rapid, sensitive methods for identification of B. mallei and B. pseudomallei are urgently needed in the interests of patient treatment and epidemiological surveillance.

METHODS

Signature sequences for sensitive, specific detection of pathogenic Burkholderia based on published genomes were identified and a qPCR assay was designed and validated.

RESULTS

A single-reaction quadruplex qPCR assay for the detection of pathogenic Burkholderia, which includes a marker for internal control of DNA extraction and amplification, was developed. The assay permits differentiation of B. mallei and B. pseudomallei strains, and probit analysis showed a very low detection limit. Use of a multicopy signature sequence permits detection of less than 1 genome equivalent per reaction.

CONCLUSIONS

The new assay permits rapid detection of pathogenic Burkholderia and combines enhanced sensitivity, species differentiation, and inclusion of an internal control for both DNA extraction and PCR amplification.

Authors+Show Affiliations

Laboratory for Zoonoses and Environmental Microbiology, National Institute for Public Health and the Environment (RIVM), Anthonie van Leeuwenhoeklaan 9, Bilthoven, MA, 3721, The Netherlands. ingmar.janse@rivm.nlNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

23409683

Citation

Janse, Ingmar, et al. "Multiplex qPCR for Reliable Detection and Differentiation of Burkholderia Mallei and Burkholderia Pseudomallei." BMC Infectious Diseases, vol. 13, 2013, p. 86.
Janse I, Hamidjaja RA, Hendriks AC, et al. Multiplex qPCR for reliable detection and differentiation of Burkholderia mallei and Burkholderia pseudomallei. BMC Infect Dis. 2013;13:86.
Janse, I., Hamidjaja, R. A., Hendriks, A. C., & van Rotterdam, B. J. (2013). Multiplex qPCR for reliable detection and differentiation of Burkholderia mallei and Burkholderia pseudomallei. BMC Infectious Diseases, 13, 86. https://doi.org/10.1186/1471-2334-13-86
Janse I, et al. Multiplex qPCR for Reliable Detection and Differentiation of Burkholderia Mallei and Burkholderia Pseudomallei. BMC Infect Dis. 2013 Feb 14;13:86. PubMed PMID: 23409683.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Multiplex qPCR for reliable detection and differentiation of Burkholderia mallei and Burkholderia pseudomallei. AU - Janse,Ingmar, AU - Hamidjaja,Raditijo A, AU - Hendriks,Amber C A, AU - van Rotterdam,Bart J, Y1 - 2013/02/14/ PY - 2012/10/02/received PY - 2013/02/08/accepted PY - 2013/2/16/entrez PY - 2013/2/16/pubmed PY - 2013/6/19/medline SP - 86 EP - 86 JF - BMC infectious diseases JO - BMC Infect Dis VL - 13 N2 - BACKGROUND: Burkholderia mallei and B. pseudomallei are two closely related species of highly virulent bacteria that can be difficult to detect. Pathogenic Burkholderia are endemic in many regions worldwide and cases of infection, sometimes brought by travelers from unsuspected regions, also occur elsewhere. Rapid, sensitive methods for identification of B. mallei and B. pseudomallei are urgently needed in the interests of patient treatment and epidemiological surveillance. METHODS: Signature sequences for sensitive, specific detection of pathogenic Burkholderia based on published genomes were identified and a qPCR assay was designed and validated. RESULTS: A single-reaction quadruplex qPCR assay for the detection of pathogenic Burkholderia, which includes a marker for internal control of DNA extraction and amplification, was developed. The assay permits differentiation of B. mallei and B. pseudomallei strains, and probit analysis showed a very low detection limit. Use of a multicopy signature sequence permits detection of less than 1 genome equivalent per reaction. CONCLUSIONS: The new assay permits rapid detection of pathogenic Burkholderia and combines enhanced sensitivity, species differentiation, and inclusion of an internal control for both DNA extraction and PCR amplification. SN - 1471-2334 UR - https://www.unboundmedicine.com/medline/citation/23409683/Multiplex_qPCR_for_reliable_detection_and_differentiation_of_Burkholderia_mallei_and_Burkholderia_pseudomallei_ L2 - https://bmcinfectdis.biomedcentral.com/articles/10.1186/1471-2334-13-86 DB - PRIME DP - Unbound Medicine ER -