Seventy amikacin-resistant clinical isolates of gram-negative bacteria belonging to nine genera were examined by immunoblotting and by DNA-DNA hybridisation for the presence of ACC(6′)1b enzyme, previously called AAC(6′)-4, or its encoding gene The organisms mostly had resistance profiles compatible with AAC(6′) production and were from South and North America, the Far East and Europe. Polyclonal (rabbit) anti-AAC(6′)-1b antisera and an intragenic () probe derived from the multiresistance plasmid pAZ007 were used. The gene was found to be widespread. Positive hybridisation, and immunologically cross-reactive proteins, were observed in 44% of the isolates examined. They were present most frequently (£70%) in isolates of and spp., but less often (£25%) in and spp. The strains that reacted with the probe produced enzymes that varied in their apparent mol. wts between 24000 and 26000. The existence of multiple electrophoretic forms of amikacin-acetylating enzymes of the ACC(6′)-1b type may be useful in epidemiological surveys of AAC(6′)-mediated amikacin resistance.


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