This Article
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 Meier, A
Right arrow Articles by Böttger, E C
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Meier, A
Right arrow Articles by Böttger, E C

 Previous Article  |  Next Article 

J Clin Microbiol. 1993 March; 31(3): 646-652

Elimination of contaminating DNA within polymerase chain reaction reagents: implications for a general approach to detection of uncultured pathogens.

A Meier, D H Persing, M Finken and E C Böttger

Institut für Medizinische Mikrobiologie, Medizinische Hochschule Hannover, Germany.

ABSTRACT

Analysis based on comparisons of 16S rRNA sequences provides a rapid and reliable approach to identifying human pathogens. By directing oligonucleotide primers at sequences conserved throughout the eubacterial kingdom, bacterial 16S ribosomal DNA sequences of virtually any member of the eubacterial kingdom can be amplified by polymerase chain reaction and subsequently analyzed by sequence determination. Indeed, automated systems for broad-range amplification, sequencing, and data analysis are now feasible and may form the basis of the next generation of automated microbial identification systems. However, identification of pathogens by this strategy is hampered by the frequent contamination of reagents used for the amplification reaction, in particular Taq polymerase, with exogenous bacterial DNA. Here, we describe detailed investigations on the use of 8-methoxypsoralen and long-wave UV light to eliminate contaminating DNA in polymerase chain reaction reagents. The clinical utility of the developed procedure was demonstrated in a case of paucibacillary osteomyelitis, for which no specific bacterial agent had been cultured.

J Clin Microbiol. 1993 March; 31(3): 646-652




This article has been cited by other articles:

  • Cheng, J.-C., Huang, C.-L., Lin, C.-C., Chen, C.-C., Chang, Y.-C., Chang, S.-S., Tseng, C.-P. (2006). Rapid Detection and Identification of Clinically Important Bacteria by High-Resolution Melting Analysis after Broad-Range Ribosomal RNA Real-Time PCR. Clin. Chem. 52: 1997-2004 [Abstract] [Full Text]  
  • Bauer, T. W., Parvizi, J., Kobayashi, N., Krebs, V. (2006). Diagnosis of Periprosthetic Infection. JBJS 88: 869-882 [Abstract] [Full Text]  
  • Dreier, J., Stormer, M., Kleesiek, K. (2004). Two Novel Real-Time Reverse Transcriptase PCR Assays for Rapid Detection of Bacterial Contamination in Platelet Concentrates. J. Clin. Microbiol. 42: 4759-4764 [Abstract] [Full Text]  
  • Newsome, T., Li, B.-J., Zou, N., Lo, S.-C. (2004). Presence of Bacterial Phage-Like DNA Sequences in Commercial Taq DNA Polymerase Reagents. J. Clin. Microbiol. 42: 2264-2267 [Abstract] [Full Text]  
  • Harris, K. A., Hartley, J. C. (2003). Development of broad-range 16S rDNA PCR for use in the routine diagnostic clinical microbiology service. J Med Microbiol 52: 685-691 [Abstract] [Full Text]  
  • Heininger, A., Binder, M., Ellinger, A., Botzenhart, K., Unertl, K., Doring, G. (2003). DNase Pretreatment of Master Mix Reagents Improves the Validity of Universal 16S rRNA Gene PCR Results. J. Clin. Microbiol. 41: 1763-1765 [Abstract] [Full Text]  
  • Gale, J. M., Romero, C. P., Tafoya, G. B., Conia, J. (2003). Application of Optical Trapping for Cells Grown on Plates: Optimization of PCR and Fidelity of DNA Sequencing of p53 Gene from a Single Cell. Clin. Chem. 49: 415-424 [Abstract] [Full Text]  
  • Grijalva, M, Horvath, R, Dendis, M, Erny, J, Benedik, J (2003). Molecular diagnosis of culture negative infective endocarditis: clinical validation in a group of surgically treated patients. Heart 89: 263-268 [Abstract] [Full Text]  
  • Tseng, C.-P., Cheng, J.-C., Tseng, C.-C., Wang, C., Chen, Y.-L., Chiu, D. T.-Y., Liao, H.-C., Chang, S.-S. (2003). Broad-Range Ribosomal RNA Real-Time PCR after Removal of DNA from Reagents: Melting Profiles for Clinically Important Bacteria. Clin. Chem. 49: 306-309 [Full Text]  
  • Millar, B. C., Xu, J., Moore, J. E. (2002). Risk Assessment Models and Contamination Management: Implications for Broad-Range Ribosomal DNA PCR as a Diagnostic Tool in Medical Bacteriology. J. Clin. Microbiol. 40: 1575-1580 [Full Text]  
  • Tay, B. K-B, Deckey, J., Hu, S. S. (2002). Spinal Infections. J Am Acad Orthop Surg 10: 188-197 [Abstract] [Full Text]  
  • Kawai, M., Matsutera, E., Kanda, H., Yamaguchi, N., Tani, K., Nasu, M. (2002). 16S Ribosomal DNA-Based Analysis of Bacterial Diversity in Purified Water Used in Pharmaceutical Manufacturing Processes by PCR and Denaturing Gradient Gel Electrophoresis. Appl. Environ. Microbiol. 68: 699-704 [Abstract] [Full Text]  
  • Jalava, J, Skurnik, M, Toivanen, A, Toivanen, P, Eerola, E (2001). Bacterial PCR in the diagnosis of joint infection. Ann Rheum Dis 60: 287-289 [Abstract] [Full Text]  
  • Han, S.-R., Zschausch, H.-C. E., Meyer, H.-G. W., Schneider, T., Loos, M., Bhakdi, S., Maeurer, M. J. (2000). Helicobacter pylori: Clonal Population Structure and Restricted Transmission within Families Revealed by Molecular Typing. J. Clin. Microbiol. 38: 3646-3651 [Abstract] [Full Text]  
  • Okhravi, N., Adamson, P., Carroll, N., Dunlop, A., Matheson, M. M., Towler, H. M. A., Lightman, S. (2000). PCR-Based Evidence of Bacterial Involvement in Eyes with Suspected Intraocular Infection. IOVS 41: 3474-3479 [Abstract] [Full Text]  
  • BECKER, K., BADEHORN, D., DEIWICK, S., PETERS, G., FEGELER, W. (2000). Molecular genotyping of Candida species with special respect to Candida (Torulopsis) glabrata strains by arbitrarily primed PCR. J Med Microbiol 49: 575-581 [Abstract] [Full Text]  
  • Corless, C. E., Guiver, M., Borrow, R., Edwards-Jones, V., Kaczmarski, E. B., Fox, A. J. (2000). Contamination and Sensitivity Issues with a Real-Time Universal 16S rRNA PCR. J. Clin. Microbiol. 38: 1747-1752 [Abstract] [Full Text]  
  • Tunney, M. M., Patrick, S., Curran, M. D., Ramage, G., Hanna, D., Nixon, J. R., Gorman, S. P., Davis, R. I., Anderson, N. (1999). Detection of Prosthetic Hip Infection at Revision Arthroplasty by Immunofluorescence Microscopy and PCR Amplification of the Bacterial 16S rRNA Gene. J. Clin. Microbiol. 37: 3281-3290 [Abstract] [Full Text]  
  • Carroll, N. M., Adamson, P., Okhravi, N. (1999). Elimination of Bacterial DNA from Taq DNA Polymerases by Restriction Endonuclease Digestion. J. Clin. Microbiol. 37: 3402-3404 [Abstract] [Full Text]  
  • Klausegger, A., Hell, M., Berger, A., Zinober, K., Baier, S., Jones, N., Sperl, W., Kofler, B. (1999). Gram Type-Specific Broad-Range PCR Amplification for Rapid Detection of 62 Pathogenic Bacteria. J. Clin. Microbiol. 37: 464-466 [Abstract] [Full Text]  
  • Hubbard, A. L., Harrison, D. J., Moyes, C., McOrist, S. (1998). Direct Detection of eae-Positive Bacteria in Human and Veterinary Colorectal Specimens by PCR. J. Clin. Microbiol. 36: 2326-2330 [Abstract] [Full Text]  
  • Gibson, N. J., Gillard, H. L., Whitcombe, D., Ferrie, R. M., Newton, C. R., Little, S. (1997). A homogeneous method for genotyping with fluorescence polarization. Clin. Chem. 43: 1336-1341 [Abstract] [Full Text]  
  • Niederhauser, C, Hofelein, C, Wegmuller, B, Luthy, J, Candrian, U (1994). Reliability of PCR decontamination systems.. Genome Res 4: 117-123 [Abstract]  
  • Cone, R W, Fairfax, M R (1993). Protocol for ultraviolet irradiation of surfaces to reduce PCR contamination.. Genome Res 3: S15-S17  


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