The Journal of Medical Microbiology, Vol 41, Issue 1 63-68, Copyright © 1994 by Society for General Microbiology

JOURNAL ARTICLE

Genomic DNA fingerprinting of clinical Haemophilus influenzae isolates by polymerase chain reaction amplification: comparison with major outer-membrane protein and restriction fragment length polymorphism analysis

A. van Belkum, B. Duim, A. Regelink, L. Moller, W. Quint and L. van Alphen
Diagnostisch Centrum SSDZ, Department of Molecular Biology, Delft, The Netherlands.

Non-capsulate strains of Haemophilus influenzae were genotyped by analysis of variable DNA segments obtained by amplification of genomic DNA with the polymerase chain reaction (PCR fingerprinting). Discrete fragments of 100-2000 bp were obtained. The reproducibility of the procedure was assessed by comparing: (i) the fingerprints of 16 colonies of a single H. influenzae strain; (ii) isolates obtained from individual sputum samples (a total of 57 H. influenzae isolates from three cystic fibrosis patients); and (iii) 17 isolates collected during an outbreak of H. influenzae infection in a local pulmonary rehabilitation centre. The discriminatory power of the method was demonstrated by showing that the PCR fingerprints of eight unrelated H. influenzae strains from sputum samples of patients with chronic obstructive pulmonary disease (COPD) and 32 strains from cystic fibrosis patients were all different. These 40 isolates also differed with respect to their restriction fragment length polymorphisms (RFLP) and major outer-membrane protein (MOMP) composition. Twelve MOMP antigenic strain variants from sputum samples of five COPD patients had identical PCR fingerprints and RFLPs. It was concluded that PCR fingerprinting is a reliable and reproducible method for genotyping non-capsulate strains of H. influenzae. The discriminatory power of PCR fingerprinting was similar to that of RFLP analysis, but the results of PCR fingerprinting were easier to interpret.

This article has been cited by other articles:

				M. E. Watson Jr, J. L. Burns, and A. L. Smith Hypermutable Haemophilus influenzae with mutations in mutS are found in cystic fibrosis sputum Microbiology, September 1, 2004; 150(9): 2947 - 2958. [Abstract] [Full Text] [PDF]

				J. R. Gilsdorf, C. F. Marrs, and B. Foxman Haemophilus influenzae: Genetic Variability and Natural Selection To Identify Virulence Factors Infect. Immun., May 1, 2004; 72(5): 2457 - 2461. [Full Text] [PDF]

				F. Roman, R. Canton, M. Perez-Vazquez, F. Baquero, and J. Campos Dynamics of Long-Term Colonization of Respiratory Tract by Haemophilus influenzae in Cystic Fibrosis Patients Shows a Marked Increase in Hypermutable Strains J. Clin. Microbiol., April 1, 2004; 42(4): 1450 - 1459. [Abstract] [Full Text] [PDF]

				E. Augustynowicz, A. Gzyl, L. Szenborn, D. Banys, G. Gniadek, and J. Slusarczyk Comparison of usefulness of randomly amplified polymorphic DNA and amplified-fragment length polymorphism techniques in epidemiological studies on nasopharyngeal carriage of non-typable Haemophilus influenzae J. Med. Microbiol., November 1, 2003; 52(11): 1005 - 1014. [Abstract] [Full Text] [PDF]

				G. Bruant, S. Watt, R. Quentin, and A. Rosenau Typing of Nonencapsulated Haemophilus Strains by Repetitive-Element Sequence-Based PCR Using Intergenic Dyad Sequences J. Clin. Microbiol., August 1, 2003; 41(8): 3473 - 3480. [Abstract] [Full Text] [PDF]

				A. Kureishi, J. V. Hirschmann, P. Bresser, L. van Alphen, R. Lutter, S. Sethi, T. F. Murphy, and N. R. Anthonisen New Strains of Bacteria and Exacerbations of COPD N. Engl. J. Med., December 19, 2002; 347(25): 2077 - 2079. [Full Text] [PDF]

				A. J. Leach, P. S. Morris, H. Smith-Vaughan, and J. D. Mathews In Vivo Penicillin MIC Drift to Extremely High Resistance in Serotype 14 Streptococcus pneumoniae Persistently Colonizing the Nasopharynx of an Infant with Chronic Suppurative Lung Disease: a Case Study Antimicrob. Agents Chemother., November 1, 2002; 46(11): 3648 - 3649. [Abstract] [Full Text] [PDF]

				M. M. Pettigrew, B. Foxman, Z. Ecevit, C. F. Marrs, and J. Gilsdorf Use of Pulsed-Field Gel Electrophoresis, Enterobacterial Repetitive Intergenic Consensus Typing, and Automated Ribotyping To Assess Genomic Variability among Strains of Nontypeable Haemophilus influenzae J. Clin. Microbiol., February 1, 2002; 40(2): 660 - 662. [Abstract] [Full Text] [PDF]

				S. Sethi and T. F. Murphy Bacterial Infection in Chronic Obstructive Pulmonary Disease in 2000: a State-of-the-Art Review Clin. Microbiol. Rev., April 1, 2001; 14(2): 336 - 363. [Abstract] [Full Text] [PDF]

				P. Gomez-De-Leon, J. I. Santos, J. Caballero, D. Gomez, L. E. Espinosa, I. Moreno, D. Piñero, and A. Cravioto Genomic Variability of Haemophilus influenzae Isolated from Mexican Children Determined by Using Enterobacterial Repetitive Intergenic Consensus Sequences and PCR J. Clin. Microbiol., July 1, 2000; 38(7): 2504 - 2511. [Abstract] [Full Text]

				A. G. Regelink, D. Dahan, L. V. M. Möller, J. W. Coulton, P. Eijk, P. Van Ulsen, J. Dankert, and L. Van Alphen Variation in the Composition and Pore Function of Major Outer Membrane Pore Protein P2 of Haemophilus influenzae from Cystic Fibrosis Patients Antimicrob. Agents Chemother., February 1, 1999; 43(2): 226 - 232. [Abstract] [Full Text]

				H. C. Smith-Vaughan, K. S. Sriprakash, A. J. Leach, J. D. Mathews, and D. J. Kemp Low Genetic Diversity of Haemophilus influenzae Type b Compared to Nonencapsulated H. influenzae in a Population in Which H. influenzae Is Highly Endemic Infect. Immun., July 1, 1998; 66(7): 3403 - 3409. [Abstract] [Full Text] [PDF]

				A. van Belkum, S. Scherer, L. van Alphen, and H. Verbrugh Short-Sequence DNA Repeats in Prokaryotic Genomes Microbiol. Mol. Biol. Rev., June 1, 1998; 62(2): 275 - 293. [Abstract] [Full Text] [PDF]

				A. R. Foxwell, J. M. Kyd, and A. W. Cripps Nontypeable Haemophilus influenzae: Pathogenesis and Prevention Microbiol. Mol. Biol. Rev., June 1, 1998; 62(2): 294 - 308. [Abstract] [Full Text] [PDF]

				L. V. M. Möller, A. G. Regelink, H. Grasselier, L. van Alphen, and J. Dankert Antimicrobial Susceptibility of Haemophilus influenzae in the Respiratory Tracts of Patients with Cystic Fibrosis Antimicrob. Agents Chemother., February 1, 1998; 42(2): 319 - 324. [Abstract] [Full Text]

				F. Geluk, P. P. Eijk, S. M. van Ham, H. M. Jansen, and L. van Alphen The Fimbria Gene Cluster of Nonencapsulated Haemophilus influenzae Infect. Immun., February 1, 1998; 66(2): 406 - 417. [Abstract] [Full Text] [PDF]