Characterisation of penA and tetM resistance genes of Neisseria gonorrhoeae isolated in southern Africa--epidemiological monitoring and resistance development.

OBJECTIVE

To investigate penA and tetM resistance gene variation of Neisseria gonorrhoeae in order to define gene types for epidemiological monitoring and resistance development.

DESIGN

Isolates of N. gonorrhoeae which were susceptible and resistant to penicillin and/or tetracycline were selected. Strains comprised South African isolates (22 from Bloemfontein, 13 from Transvaal, 20 from the Cape) and 15 Botswana and 4 Namibia isolates. The penA genes (2 kb) of all strains and tetM genes (765 bp) of 11 high-level tetracycline-resistant strains were amplified and restricted with HpaII.

RESULTS AND CONCLUSIONS

Twelve different HpaII fingerprint patterns were obtained from the 74 isolates analysed for penicillin-binding protein (PBP) 2 gene (penA) alterations. Focusing on the transpeptidase domain, 25 isolates (3 whole gene patterns, minimal inhibitory concentrations (MICs) < or = 0.03-0.125 micrograms/ml) had restriction sites equivalent to those previously described for a susceptible strain. Of the remaining 9 PBP 2 gene groups, 25 strains fell into a designated group E. Penicillin/ penicillin + clavulanic acid MICs determined on these group E isolates gave a range of 0.125-2.0 micrograms/ml, although MICs against 4 strains were < or = 0.03 micrograms/ml. MICs of penicillin/penicillin + clavulanic acid for the 24 isolates that contained altered PBP 2 transpeptidase gene regions not designated group E were only < or = 0.03-0.125 micrograms/ml. The lack of a HpaII restriction site at nucleotide 1934 in the PBP 2 gene of group E strains was indicative of a small terminal region of N. cinerea DNA. This gene block, which was found in all the southern African areas studied, appears to predispose isolates to increased penicillin resistance. The 25.2 MDa conjugative plasmid carrying the tetM resistance determinant was readily demonstrated in 11 Botswana/Namibia isolates exhibiting high-level resistance to tetracycline (MICs > or = 16 micrograms/ml). The tetM gene was shown to be of the American type.