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

Abstract

ABSTRACT 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 Sma I 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.