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Journal of Clinical Microbiology, April 2004, p. 1727-1730, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1727-1730.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Identification of Bacteroides thetaiotaomicron on the Basis of an Unexpected Specific Amplicon of Universal 16S Ribosomal DNA PCR

Lee-Jene Teng,1,2 Po-Ren Hsueh,1 Yu-Hsuan Huang,2 and Jui-Chang Tsai3,4*

Department of Laboratory Medicine,1 Division of Neurosurgery, Department of Surgery, National Taiwan University Hospital,3 School of Medical Technology,2 Center for Optoelectronic Biomedicine, National Taiwan University College of Medicine, Taipei, Taiwan4

Received 31 August 2003/ Returned for modification 24 October 2003/ Accepted 18 December 2003

We applied a set of commonly used universal primers (primers RW01 and DG74) to amplify partial fragments of 16S ribosomal DNA for bacterial identification and found an unexpected amplicon (547 bp), in addition to the expected 362-bp product, in samples containing Bacteroides thetaiotaomicron. It was demonstrated that the internal sequence (508 bp, excluding the primers) of the 547-bp amplicon was identical to the genomic sequence from nucleotide positions 165800 to 166307 of B. thetaiotaomicron type strain VPI-5482 by a BLAST search of the sequences in the GenBank database. The existence of this unexpected yet specific amplicon strongly indicated the presence of B. thetaiotaomicron in the sample, and it was found that it could be used to discriminate B. thetaiotaomicron from closely related species. Another set of PCR primers specific for B. thetaiotaomicron was developed on the basis of the sequence of this 547-bp genomic fragment. Both PCR-based assays showed the same sensitivity (88%) and specificity (100%).

* Corresponding author. Mailing address: Division of Neurosurgery, Department of Surgery, National Taiwan University College of Medicine, Rm. 824, 8th Floor, No. 7, Chung-Shan S. Rd., Taipei 100, Taiwan. Phone: 886-2-23123456, ext. 5422. Fax: 886-2-23582009. E-mail: jctsai{at}ha.mc.ntu.edu.tw.

Journal of Clinical Microbiology, April 2004, p. 1727-1730, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1727-1730.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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