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Journal of Clinical Microbiology, May 1999, p. 1524-1531, Vol. 37, No. 5
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Pulsed-Field Gel Electrophoresis Used To Investigate Genetic Diversity of Haemophilus influenzae Type b Isolates in Australia Shows Differences between Aboriginal and Non-Aboriginal Isolates

Patricia Ezekiel Moor,1,* Peter C. Collignon,2 and Gwendolyn L. Gilbert3

Division of Biochemistry and Molecular Biology, Australian National University, Australian Capital Territory 0200,1 Infectious Disease Unit, Canberra Hospital, Woden 2606,2 and Centre for Infectious Diseases and Microbiology, Institute of Clinical Pathology and Medical Research, Westmead Hospital, Westmead 2145,3 Australia

Received 13 October 1998/Returned for modification 8 December 1998/Accepted 28 January 1999

We used pulsed-field gel electrophoresis to study the epidemiology and population structure of Haemophilus influenzae type b. DNAs from 187 isolates recovered between 1985 and 1993 from Aboriginal children (n = 76), non-Aboriginal children (n = 106), and non-Aboriginal adults (n = 5) in urban and rural regions across Australia were digested with the SmaI restriction endonuclease. Patterns of 13 to 17 well-resolved fragments (size range, ~8 to 500 kb) defining 67 restriction fragment length polymorphism (RFLP) types were found. Two types predominated. One type (n = 37) accounted for 35 (46%) of the isolates from Aboriginals and 2 (2%) of the isolates from non-Aboriginals, and the other type (n = 41) accounted for 2 (3%) of the isolates from Aboriginals and 39 (35%) of the isolates from non-Aboriginals. Clustering revealed seven groups at a genetic distance of ~50% similarity in a tree-like dendrogram. They included two highly divergent groups representing 50 (66%) isolates from Aboriginals and 6 (5%) isolates from non-Aboriginals and another genetically distinct group representing 7 (9%) isolates from Aboriginals and 81 (73%) isolates from non-Aboriginals. The results showed a heterogeneous clonal population structure, with the isolates of two types accounting for 42% of the sample. There was no association between RFLP type and the diagnosis of meningitis or epiglottitis, age, sex, date of collection, or geographic location, but there was a strong association between the origin of isolates from Aboriginal children and RFLP type F2a and the origin of isolates from non-Aboriginal children and RFLP type A8b. The methodology discriminated well among the isolates (D = 0.91) and will be useful for the monitoring of postvaccine isolates of H. influenzae type b.

* Corresponding author. Mailing address: Gadi Research Centre, Faculty of Applied Science, University of Canberra, Canberra, ACT 2601, Australia. Phone: 61-6-6201-5451. Fax: 61-6-6201-5727. E-mail: patmoor{at}ozemail.com.au.

Journal of Clinical Microbiology, May 1999, p. 1524-1531, Vol. 37, No. 5
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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