1887

Abstract

A total of 67 field isolates from the USA, Israel and Australia, and 10 reference strains, were characterized by gene-targeted sequencing (GTS) analysis of portions of the putative cytadhesin gene, the cytadhesin gene, the cytadhesin gene, and an uncharacterized hypothetical surface lipoprotein-encoding gene designated genome coding DNA sequence (CDS) MGA_0319. The regions of the surface-protein-encoding genes targeted in this analysis were found to be stable within a strain, after sequencing different passages of reference strains. Gene sequences were first analysed on the basis of gene size polymorphism. The and genes are characterized by the presence of different nucleotide insertions/deletions. However, differentiation of isolates based solely on / PCR size polymorphism was not found to be a reliable method to differentiate among isolates. On the other hand, GTS analysis based on the nucleotide sequence identities of individual and multiple genes correlated with epidemiologically linked isolates and with random amplified polymorphic DNA (RAPD) analysis. GTS analysis of individual genes, , MGA_0319, and , identified 17, 16, 20 and 22 sequence types, respectively. GTS analysis using multiple gene sequences / and /MGA_0319// identified 38 and 40 sequence types, respectively. GTS of multiple surface-protein-encoding genes showed better discriminatory power than RAPD analysis, which identified 36 pattern types from the same panel of strains. These results are believed to provide the first evidence that typing of isolates by GTS analysis of surface-protein genes is a sensitive and reproducible typing method and will allow rapid global comparisons between laboratories.

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2005-06-01
2019-10-17
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vol. , part 6, pp. 1883 - 1893

Dendrograms: 1- , 2-MGA_0319, 3- , 4- , 5- / , 6- /MGA_0319/ / . Dendrograms were constructed from 77 sequences. Dendrograms 1 to 4 were constructed from individual gene sequences, and dendrograms 5 and 6 were constructed with multiple gene sequences. Dendrograms were generated using the neighbour joining (NJ) method with 1000 bootstraps replicates using MEGA2 ( www.megasoftware.net). The different sequence clusters within the dendrogram are indicated in roman numbers. Next to each isolate the RAPD type is indicated in parentheses, and the isolate sequence type within the cluster is indicated by a lower-case letter. [ PDF file] (570 KB)



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