Association between dnd gene cluster and the DNA degradation phenotype of untypeable Escherichia coli strains
Pulsed-field gel electrophoresis (PFGE) is generally accepted as the “gold standard” subtyping method for Escherichia coli. However, strains suffering from in-gel DNA degradation (probably due to presence of dnd operon) during PFGE run can impact on the molecular epidemiological data. Although DNA...
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| 格式: | Conference or Workshop Item |
| 语言: | English |
| 出版: |
2014
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| 主题: | |
| 在线阅读: | http://eprints.unisza.edu.my/452/1/FH03-FPSK-14-01954.pdf http://eprints.unisza.edu.my/452/ |
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| 总结: | Pulsed-field gel electrophoresis (PFGE) is generally accepted as the “gold standard” subtyping method for
Escherichia coli. However, strains suffering from in-gel DNA degradation (probably due to presence of dnd
operon) during PFGE run can impact on the molecular epidemiological data. Although DNA degradation is
common in many different bacteria, studies reporting the association of the Dnd phenotype and genotype are
still lacking. The objective of this study was thus to improve the typeability of E. coli and to determine the
association between the presence of dnd gene cluster and the Dnd phenotype. We would also aim to determine
whether the Dnd phenotype is associated with an inherent clonal trait. A total of 60 human and zoonotic E. coli
strains including 12 strains with the Dnd phenotype (Dnd+) were studied. PCR detection of dndB-E genes of
the 60 strains was carried out. PFGE was performed on XbaI-digested genomic DNA of Dnd+ strains with and
without the addition of thiourea. There was a 100% improvement on the typeability of Dnd+ E. coli strains. Dnd
phenotype was also observed in both clonal and genetically diverse E. coli strains. Nine out of the 12 Dnd+
E. coli strains shared indistinguishable PFGE profile. Another three Dnd+ strains were genetically diverse. The
dndBCDE genes (dnd operon) were detected in all Dnd+ E. coli strains by PCR and association between the phenotype and genotype of DNA degradation was also found. The untypeable problem of Dnd+ strains should be
overcome as all of the Dnd+ strains in this study were obtained from unhealthy hosts and their epidemiological
data may be of clinical importance. |
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