Wystepowanie wybranych genow loci waa i wb zwiazanych z wytwarzaniem lipopolisacharydu u referencyjnych i izolowanych z materialu klinicznego szczepow paleczek Klebsiella pneumoniae

Określono częstość występowania 11 wybranych genów loci waa i wb związanych z biosyntezą oligocukru rdzenia lipopolisacharydu (LPS) i antygenu O pałeczek Klebsiella. Zbadano 26 wybranych szczepów wzorcowych dla antygenu K reprezentujących wyróżniane obecnie grupy serologiczne pałeczek K. pneumoniae a także 19 szczepów epidemicznych i izolowanych od przypadkowych osób na terenie kraju. Zróżnicowanie częstości występowania poszukiwanych genów w grupie badanych szczepów stanowiło podstawę wyodrębnienia 21 genotypów. Wykazano przydatność genów z loci waa i wb do genotypowania pałeczek K. pneumoniae.

The goal of presented study was to determine by PCR differences in existence or homology level of selected genes involved in A', pneumoniae lipopolysaccharide (LPS) synthesis and application of obtained results for genotyping. Number of 26 reference strains of K. pneumoniae belonging to sero- groups 01,02a, 02a2e, 02a2e2h, 02a2f2g, 03,04,05,07,08, and 012 was tested together with 13 epidemic strains from 5 outbreaks and б casual isolates for the existence of 7 (waaA, waaE, waaL, waaQ, waaZ, waaX and uge) and 4 (wbdA, wbdC, manB, wbbO) genes of the waa and wb clusters for LPS biosynthesis. Based on PCR results, 10 and 11 genotypes were distinguished in tested strains for genes from waa and wb clusters respectively. Derived dendrograms were topologically dissimilar, however observed correlation between clonal groups and O-group was marginal for both compared clusters. Since we aimed to develop genotyping method for K. pneumoniae, genes from clusters waa and wb were used together to enhance the distinguishing capacity. Twenty-one genotypes were distinguished in 45 tested strains (DI=0,46) when 11 genes were applied for typing. Although no apparent correlation between genotype and serogroup was observed, epidemic isolates from 5 outbreaks were diversified into 5 genotypes, whereas strains from the same outbreak were indistinguishable. Described here genotyping method is determinative and was found time and cost effective. This method may be applied in every clinical laboratory equipped in an ordinary PCR apparatus.