Accurate identification of fastidious Gram-negative rods: integration of both conventional phenotypic methods and 16S rRNA gene analysis

BMC Microbiology

Methodology Article

Andrea Zbinden¹  Reinhard Zbinden¹  Guido V Bloemberg¹  Susanne Abels²  Maria G de Melo Oliveira³ 

[1] Institute of Medical Microbiology, University of Zurich, 8006, Zurich, Switzerland;Institute of Medical Microbiology, University of Zurich, 8006, Zurich, Switzerland;Institute for Hygiene and Public Health, University Hospital, 53105, Bonn, Germany;Institute of Medical Microbiology, University of Zurich, 8006, Zurich, Switzerland;Policlínica Nascente, Prefeitura Municipal de Fortaleza, Fortalez-Ceará, Brasil;

关键词: Fastidious Gram-negative rods;    16S rRNA gene;    Conventional phenotypic methods;   

DOI  :  10.1186/1471-2180-13-162

 received in 2013-01-05, accepted in 2013-07-12,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundAccurate identification of fastidious Gram-negative rods (GNR) by conventional phenotypic characteristics is a challenge for diagnostic microbiology. The aim of this study was to evaluate the use of molecular methods, e.g., 16S rRNA gene sequence analysis for identification of fastidious GNR in the clinical microbiology laboratory.ResultsA total of 158 clinical isolates covering 20 genera and 50 species isolated from 1993 to 2010 were analyzed by comparing biochemical and 16S rRNA gene sequence analysis based identification. 16S rRNA gene homology analysis identified 148/158 (94%) of the isolates to species level, 9/158 (5%) to genus and 1/158 (1%) to family level. Compared to 16S rRNA gene sequencing as reference method, phenotypic identification correctly identified 64/158 (40%) isolates to species level, mainly Aggregatibacter aphrophilus, Cardiobacterium hominis, Eikenella corrodens, Pasteurella multocida, and 21/158 (13%) isolates correctly to genus level, notably Capnocytophaga sp.; 73/158 (47%) of the isolates were not identified or misidentified.ConclusionsWe herein propose an efficient strategy for accurate identification of fastidious GNR in the clinical microbiology laboratory by integrating both conventional phenotypic methods and 16S rRNA gene sequence analysis. We conclude that 16S rRNA gene sequencing is an effective means for identification of fastidious GNR, which are not readily identified by conventional phenotypic methods.

【 授权许可】

Unknown   
© de Melo Oliveira et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.