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

BMC Microbiology

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

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

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

Others  :  1143422
DOI  :  10.1186/1471-2180-13-162

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

【 摘 要 】

Background

Accurate 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.

Results

A 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.

Conclusions

We 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】

   
2013 de Melo Oliveira et al.; licensee BioMed Central Ltd.

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