This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Fontana, C.
Right arrow Articles by Favalli, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Fontana, C.
Right arrow Articles by Favalli, C.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, February 2005, p. 615-619, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.615-619.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Use of the MicroSeq 500 16S rRNA Gene-Based Sequencing for Identification of Bacterial Isolates That Commercial Automated Systems Failed To Identify Correctly

Carla Fontana,1,2* Marco Favaro,1 Marco Pelliccioni,2 Enrico Salvatore Pistoia,1 and Cartesio Favalli1,2

Department of Experimental Medicine and Biochemical Sciences, "Tor Vergata" University of Rome,1 Clinical Microbiology Laboratories, Policlinic of Tor Vergata, Rome, Italy2

Received 13 May 2004/ Returned for modification 4 August 2004/ Accepted 24 September 2004

Reliable automated identification and susceptibility testing of clinically relevant bacteria is an essential routine for microbiology laboratories, thus improving patient care. Examples of automated identification systems include the Phoenix (Becton Dickinson) and the VITEK 2 (bioMérieux). However, more and more frequently, microbiologists must isolate "difficult" strains that automated systems often fail to identify. An alternative approach could be the genetic identification of isolates; this is based on 16S rRNA gene sequencing and analysis. The aim of the present study was to evaluate the possible use of MicroSeq 500 (Applera) for sequencing the 16S rRNA gene to identify isolates whose identification is unobtainable by conventional systems. We analyzed 83 "difficult" clinical isolates: 25 gram-positive and 58 gram-negative strains that were contemporaneously identified by both systems—VITEK 2 and Phoenix—while genetic identification was performed by using the MicroSeq 500 system. The results showed that phenotypic identifications by VITEK 2 and Phoenix were remarkably similar: 74% for gram-negative strains (43 of 58) and 80% for gram-positive strains were concordant by both systems and also concordant with genetic characterization. The exceptions were the 15 gram-negative and 9 gram-positive isolates whose phenotypic identifications were contrasting or inconclusive. For these, the use of MicroSeq 500 was fundamental to achieving species identification. In clinical microbiology the use of MicroSeq 500, particularly for strains with ambiguous biochemical profiles (including slow-growing strains), identifies strains more easily than do conventional systems. Moreover, MicroSeq 500 is easy to use and cost-effective, making it applicable also in the clinical laboratory.

* Corresponding author. Mailing address: Department of Experimental Medicine and Biochemical Sciences, "Tor Vergata" University of Rome, Via Montpellier 1, 00133 Rome, Italy. Phone: 039-6-20902158. Fax: 039-6-20902078. E-mail: carla.fontana{at}uniroma2.it.

Journal of Clinical Microbiology, February 2005, p. 615-619, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.615-619.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Woo, P. C. Y., Teng, J. L. L., Wu, J. K. L., Leung, F. P. S., Tse, H., Fung, A. M. Y., Lau, S. K. P., Yuen, K.-y. (2009). Guidelines for interpretation of 16S rRNA gene sequence-based results for identification of medically important aerobic Gram-positive bacteria. J Med Microbiol 58: 1030-1036 [Abstract] [Full Text]  
  • Leader, B. T., Frye, J. G., Hu, J., Fedorka-Cray, P. J., Boyle, D. S. (2009). High-Throughput Molecular Determination of Salmonella enterica Serovars by Use of Multiplex PCR and Capillary Electrophoresis Analysis. J. Clin. Microbiol. 47: 1290-1299 [Abstract] [Full Text]  
  • Song, S. H., Park, J. S., Kwon, H. R., Kim, S. H., Kim, H. B., Chang, H. E., Park, K. U., Song, J., Kim, E. C. (2009). Human bloodstream infection caused by Staphylococcus pettenkoferi. J Med Microbiol 58: 270-272 [Abstract] [Full Text]  
  • Janda, J. M., Abbott, S. L. (2007). 16S rRNA Gene Sequencing for Bacterial Identification in the Diagnostic Laboratory: Pluses, Perils, and Pitfalls. J. Clin. Microbiol. 45: 2761-2764 [Full Text]  
  • Woo, P. C Y, Chung, L. M W, Teng, J. L L, Tse, H., Pang, S. S Y, Lau, V. Y T, Wong, V. W K, Kam, K.-l., Lau, S. K P, Yuen, K.-Y. (2007). In silico analysis of 16S ribosomal RNA gene sequencing-based methods for identification of medically important anaerobic bacteria. J. Clin. Pathol. 60: 576-579 [Abstract] [Full Text]  
  • Loiez, C., Wallet, F., Pischedda, P., Renaux, E., Senneville, E., Mehdi, N., Courcol, R. J. (2007). First Case of Osteomyelitis Caused by "Staphylococcus pettenkoferi". J. Clin. Microbiol. 45: 1069-1071 [Abstract] [Full Text]  
  • Layer, F., Ghebremedhin, B., Moder, K.-A., Konig, W., Konig, B. (2006). Comparative study using various methods for identification of Staphylococcus species in clinical specimens.. J. Clin. Microbiol. 44: 2824-2830 [Abstract] [Full Text]  
  • Lin, J.-C., Hou, S.-J., Huang, L.-U., Sun, J.-R., Chang, W.-K., Lu, J.-J. (2006). Acute Cholecystitis Accompanied by Acute Pancreatitis Potentially Caused by Dolosigranulum pigrum.. J. Clin. Microbiol. 44: 2298-2299 [Abstract] [Full Text]  
  • Woo, P. C. Y., Tse, H., Wong, S. S. Y., Tse, C. W. S., Fung, A. M. Y., Tam, D. M. W., Lau, S. K. P., Yuen, K.-y. (2005). Life-Threatening Invasive Helcococcus kunzii Infections in Intravenous-Drug Users and ermA-Mediated Erythromycin Resistance. J. Clin. Microbiol. 43: 6205-6208 [Abstract] [Full Text]  
  • Wallet, F., Loiez, C., Renaux, E., Lemaitre, N., Courcol, R. J. (2005). Performances of VITEK 2 Colorimetric Cards for Identification of Gram-Positive and Gram-Negative Bacteria. J. Clin. Microbiol. 43: 4402-4406 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»