Multiplex qPCR for reliable detection and differentiation of Burkholderia mallei and Burkholderia pseudomallei. 2013

Ingmar Janse, and Raditijo A Hamidjaja, and Amber C A Hendriks, and Bart J van Rotterdam
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.nl

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.

UI MeSH Term Description Entries
D008554 Melioidosis A disease of humans and animals that resembles GLANDERS. It is caused by BURKHOLDERIA PSEUDOMALLEI and may range from a dormant infection to a condition that causes multiple abscesses, PNEUMONIA and BACTEREMIA. Whitmore's Disease,Burkholderia pseudomallei Infection,Burkholderia pseudomallei Infections,Disease, Whitmore's,Infection, Burkholderia pseudomallei,Melioidoses,Whitmore Disease
D005896 Glanders A contagious disease of horses that can be transmitted to humans. It is caused by BURKHOLDERIA MALLEI and characterized by ulceration of the respiratory mucosa and an eruption of nodules on the skin.
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D001426 Bacterial Proteins Proteins found in any species of bacterium. Bacterial Gene Products,Bacterial Gene Proteins,Gene Products, Bacterial,Bacterial Gene Product,Bacterial Gene Protein,Bacterial Protein,Gene Product, Bacterial,Gene Protein, Bacterial,Gene Proteins, Bacterial,Protein, Bacterial,Proteins, Bacterial
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity
D015203 Reproducibility of Results The statistical reproducibility of measurements (often in a clinical context), including the testing of instrumentation or techniques to obtain reproducible results. The concept includes reproducibility of physiological measurements, which may be used to develop rules to assess probability or prognosis, or response to a stimulus; reproducibility of occurrence of a condition; and reproducibility of experimental results. Reliability and Validity,Reliability of Result,Reproducibility Of Result,Reproducibility of Finding,Validity of Result,Validity of Results,Face Validity,Reliability (Epidemiology),Reliability of Results,Reproducibility of Findings,Test-Retest Reliability,Validity (Epidemiology),Finding Reproducibilities,Finding Reproducibility,Of Result, Reproducibility,Of Results, Reproducibility,Reliabilities, Test-Retest,Reliability, Test-Retest,Result Reliabilities,Result Reliability,Result Validities,Result Validity,Result, Reproducibility Of,Results, Reproducibility Of,Test Retest Reliability,Validity and Reliability,Validity, Face
D016957 Burkholderia pseudomallei A species of gram-negative, aerobic bacteria that causes MELIOIDOSIS. It has been isolated from soil and water in tropical regions, particularly Southeast Asia. Pseudomonas pseudomallei
D042726 Burkholderia mallei A species of gram-negative bacteria parasitic on HORSES and DONKEYS causing GLANDERS, which can be transmitted to humans. Pseudomonas mallei
D058889 Molecular Typing Using MOLECULAR BIOLOGY techniques, such as DNA SEQUENCE ANALYSIS; PULSED-FIELD GEL ELECTROPHORESIS; and DNA FINGERPRINTING, to identify, classify, and compare organisms and their subtypes. Typing, Molecular

Related Publications

Ingmar Janse, and Raditijo A Hamidjaja, and Amber C A Hendriks, and Bart J van Rotterdam
March 2005, FEMS immunology and medical microbiology,
Ingmar Janse, and Raditijo A Hamidjaja, and Amber C A Hendriks, and Bart J van Rotterdam
April 2004, Molecular and cellular probes,
Ingmar Janse, and Raditijo A Hamidjaja, and Amber C A Hendriks, and Bart J van Rotterdam
September 1998, Journal of clinical microbiology,
Ingmar Janse, and Raditijo A Hamidjaja, and Amber C A Hendriks, and Bart J van Rotterdam
June 2009, Military medicine,
Ingmar Janse, and Raditijo A Hamidjaja, and Amber C A Hendriks, and Bart J van Rotterdam
November 2006, Diagnostic microbiology and infectious disease,
Ingmar Janse, and Raditijo A Hamidjaja, and Amber C A Hendriks, and Bart J van Rotterdam
January 2009, Molecular and cellular probes,
Ingmar Janse, and Raditijo A Hamidjaja, and Amber C A Hendriks, and Bart J van Rotterdam
September 2012, Journal of microbiological methods,
Ingmar Janse, and Raditijo A Hamidjaja, and Amber C A Hendriks, and Bart J van Rotterdam
January 2002, Microbiology and immunology,
Ingmar Janse, and Raditijo A Hamidjaja, and Amber C A Hendriks, and Bart J van Rotterdam
January 2015, Analytical and bioanalytical chemistry,
Copied contents to your clipboard!