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Journal of Clinical Microbiology, December 1999, p. 3809-3814, Vol. 37, No. 12
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Molecular Epidemiological Investigation Using a Randomly Amplified Polymorphic DNA Assay of Burkholderia cepacia Isolates from Nosocomial Outbreaks

Mitsuhiro Okazaki,1,* Takashi Watanabe,2 Koji Morita,3 Yoshimi Higurashi,4 Koji Araki,1 Naoko Shukuya,1 Shigeyuki Baba,5 Noboru Watanabe,3 Teruo Egami,1 Nobushige Furuya,1 Masato Kanamori,3 Shuji Shimazaki,6 and Hidemasa Uchimura1,2

Department of Clinical Laboratory1 and Trauma and Critical Care Center,6 Kyorin University Hospital, and Department of Clinical Pathology,2 Kyorin University School of Medicine, Tokyo 181-8611, Department of Microbiology, Kyorin University School of Health Sciences, Tokyo 192-8508,3 and Department of Clinical Laboratory, University of Tokyo Hospital,4 and Department of Infection Control and Prevention,5 Tokyo University School of Medicine, Tokyo 113-0033, Japan

Received 13 January 1999/Returned for modification 27 February 1999/Accepted 21 August 1999

We experienced two Burkholderia cepacia outbreaks over a 1-year period. During this period, 28 B. cepacia isolates were obtained from clinical specimens, and 2 were obtained from environmental specimens (i.e., from a nebulizer solution and a nebulizer tube). These 30 isolates were subjected to the PCR-based randomly amplified polymorphic DNA (RAPD) assay as well as to pulsed-field gel electrophoresis (PFGE). In the first outbreak, in which eight patients hospitalized in the Trauma and Critical Care Center were involved, the RAPD assay revealed that all 20 isolates obtained from clinical specimens and the 2 isolates from environmental specimens had identical DNA profiles. These RAPD data enabled us to pinpoint a possible source and to take countermeasures to prevent further spread of the epidemic-causing strain. In the second outbreak, two consecutive B. cepacia infection/colonization cases were seen in the surgery ward. The RAPD profiles of four isolates obtained were again identical, but they were distinct from those seen in the first outbreak, clearly indicating that the second outbreak was not related to the first. Thus, our experience demonstrated that the RAPD assay is a useful and reliable tool for epidemiological studies of B. cepacia isolates from nosocomial outbreaks. Since the RAPD assay could provide discriminatory potential and reproducibility comparable to those of the widely used PFGE assay with less complexity and in a shorter time, the introduction of the RAPD assay into hospital microbiology laboratories as a routine technique may help prevent nosocomial outbreaks.

* Corresponding author. Mailing address: Department of Clinical Laboratory, Kyorin University Hospital, 6-20-2 Shinkawa, Mitaka, Tokyo 181-8611, Japan. Phone: (81) 422 47 5511, ext. 2805. Fax: (81) 422 79 3471. E-mail: ur9t-wtnb{at}asahi-net.or.jp.

Journal of Clinical Microbiology, December 1999, p. 3809-3814, Vol. 37, No. 12
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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