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Nontypeable Haemophilus influenzae Meningitis in Children: Phenotypic and Genotypic Characterization of Isolates

Cardines, Rita DBiol*; Giufrè, Maria DBiol*; Mastrantonio, Paola DBiol*; degli Atti, Marta Luisa Ciofi MD; Cerquetti, Marina DBiol*

The Pediatric Infectious Disease Journal: July 2007 - Volume 26 - Issue 7 - p 577-582
doi: 10.1097/INF.0b013e3180616715
Original Studies

Background: With the decline in the incidence of invasive Haemophilus influenzae type b disease as result of routine immunization of infants, the potential emergence of nontypeable H. influenzae (NTHi) strains as important pathogens has been suggested.

Methods: From June 1997 to July 2006, 9 cases of NTHi meningitis in children aged ≤60 months were detected. The 9 NTHi isolates were characterized. Antimicrobial susceptibility patterns were determined by E-test. The transpeptidase domain of penicillin binding protein 3 of a β-lactamase negative ampicillin-resistant strain was sequenced. Genetic relatedness among isolates was assessed by pulsed field gel electrophoresis and by multilocus sequence typing. The presence of HMW and Hia adhesins and hemagglutinating fimbriae was investigated by PCR and Western Blotting.

Results: The 9 cases of NTHi meningitis did not occur in specific risk groups, except for one patient. Of the 9 NTHi isolates, 2 were β-lactamase producers and 1 showed the β-lactamase negative ampicillin-resistant phenotype. Sequencing of the penicillin binding protein 3 revealed novel amino acid substitutions. A high degree of genetic diversity among isolates was demonstrated by pulsed field gel electrophoresis. Multilocus sequence genotyping confirmed that the 9 NTHi isolates did not belong to related phylogenetic clusters. HMW adhesins were found in 2 isolates, and 5 strains possessed Hia. No hemagglutinating fimbriae were detected, even though 2 isolates contained hifA gene sequences.

Conclusion: NTHi isolates from cases of meningitis in children are genetically diverse. Distribution of adhesins among the isolates we examined is unusual: most strains express Hia that generally occurs in a minority of strains in NTHi, suggesting that this adhesin may play a role in virulence mechanisms of NTHi causing meningitis.

From the *Department of Infectious, Parasitic and Immune-mediated Diseases; and †National Centre of Epidemiology Surveillance and Health Promotion, Istituto Superiore di Sanità, Rome, Italy.

Accepted for publication March 22, 2007.

Address for correspondence: Marina Cerquetti, DBiol, Department of Infectious, Parasitic and Immune-mediated Diseases, Istituto Superiore di Sanità, Viale Regina Elena, 299, 00161 Roma, Italy. E-mail:

© 2007 Lippincott Williams & Wilkins, Inc.