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Rapid presumptive identification of Burkholderia pseudomallei with real-time PCR assays using fluorescent hybridization probes.
Mol Cell Probes. 2005 Feb; 19(1):9-20.MC

Abstract

Burkholderia pseudomallei (the etiologic agent of melioidosis) can cause pyogenic or granulomatous lesions in almost any organ. Septicemia has a case fatality rate of >40%. Early diagnosis and appropriate antibiotic therapy are crucial for survival, but cultivation, biochemical identification, and conventional PCR of B. pseudomallei are time consuming. We established real-time PCR assays using fluorescent hybridization probes targeting the 16S rDNA, the flagellin C (fliC) and the ribosomal protein subunit S21 (rpsU) genes. The test sensitivity and specificity were assessed with a representative panel of 39 B. pseudomallei, 9 B. mallei, 126 other Burkholderia strains of 29 species, and 45 clinically relevant non-Burkholderia organisms. The detection limit for the 16S rDNA, fliC, and rpsU assay was 40, 40, and 400 genome equivalents per reaction, however, in spiked blood samples it was 300, 300, and 3000, respectively. Specificity, positive and negative predictive value of the assays was 100%. In conclusion, we recommend the use of the 16S rDNA and/or fliC real-time PCR assays for the rapid identification of B. mallei and B. pseudomallei in positive blood cultures or from suspicious bacterial colonies.

Authors+Show Affiliations

Federal Armed Forces Institute of Microbiology, Neuherbergstrasse 11, Munich 80937, Germany. herbert.tomaso@web.deNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

15652215

Citation

Tomaso, Herbert, et al. "Rapid Presumptive Identification of Burkholderia Pseudomallei With Real-time PCR Assays Using Fluorescent Hybridization Probes." Molecular and Cellular Probes, vol. 19, no. 1, 2005, pp. 9-20.
Tomaso H, Pitt TL, Landt O, et al. Rapid presumptive identification of Burkholderia pseudomallei with real-time PCR assays using fluorescent hybridization probes. Mol Cell Probes. 2005;19(1):9-20.
Tomaso, H., Pitt, T. L., Landt, O., Al Dahouk, S., Scholz, H. C., Reisinger, E. C., Sprague, L. D., Rathmann, I., & Neubauer, H. (2005). Rapid presumptive identification of Burkholderia pseudomallei with real-time PCR assays using fluorescent hybridization probes. Molecular and Cellular Probes, 19(1), 9-20.
Tomaso H, et al. Rapid Presumptive Identification of Burkholderia Pseudomallei With Real-time PCR Assays Using Fluorescent Hybridization Probes. Mol Cell Probes. 2005;19(1):9-20. PubMed PMID: 15652215.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Rapid presumptive identification of Burkholderia pseudomallei with real-time PCR assays using fluorescent hybridization probes. AU - Tomaso,Herbert, AU - Pitt,Tyrone L, AU - Landt,Olfert, AU - Al Dahouk,Sascha, AU - Scholz,Holger C, AU - Reisinger,Emil C, AU - Sprague,Lisa D, AU - Rathmann,Ilka, AU - Neubauer,Heinrich, PY - 2004/04/20/received PY - 2004/08/02/accepted PY - 2005/1/18/pubmed PY - 2005/7/1/medline PY - 2005/1/18/entrez SP - 9 EP - 20 JF - Molecular and cellular probes JO - Mol Cell Probes VL - 19 IS - 1 N2 - Burkholderia pseudomallei (the etiologic agent of melioidosis) can cause pyogenic or granulomatous lesions in almost any organ. Septicemia has a case fatality rate of >40%. Early diagnosis and appropriate antibiotic therapy are crucial for survival, but cultivation, biochemical identification, and conventional PCR of B. pseudomallei are time consuming. We established real-time PCR assays using fluorescent hybridization probes targeting the 16S rDNA, the flagellin C (fliC) and the ribosomal protein subunit S21 (rpsU) genes. The test sensitivity and specificity were assessed with a representative panel of 39 B. pseudomallei, 9 B. mallei, 126 other Burkholderia strains of 29 species, and 45 clinically relevant non-Burkholderia organisms. The detection limit for the 16S rDNA, fliC, and rpsU assay was 40, 40, and 400 genome equivalents per reaction, however, in spiked blood samples it was 300, 300, and 3000, respectively. Specificity, positive and negative predictive value of the assays was 100%. In conclusion, we recommend the use of the 16S rDNA and/or fliC real-time PCR assays for the rapid identification of B. mallei and B. pseudomallei in positive blood cultures or from suspicious bacterial colonies. SN - 0890-8508 UR - https://www.unboundmedicine.com/medline/citation/15652215/Rapid_presumptive_identification_of_Burkholderia_pseudomallei_with_real_time_PCR_assays_using_fluorescent_hybridization_probes_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0890-8508(04)00076-3 DB - PRIME DP - Unbound Medicine ER -