TY - JOUR
T1 - Escherichia coli O-antigen gene clusters of serogroups O62, O68, O131, O140, O142, and O163
T2 - DNA sequences and similarity between O62 and O68, and PCR-based serogrouping
AU - Liu, Yanhong
AU - Yan, Xianghe
AU - DebRoy, Chitrita
AU - Fratamico, Pina M.
AU - Needleman, David S.
AU - Li, Robert W.
AU - Wang, Wei
AU - Losada, Liliana
AU - Brinkac, Lauren
AU - Radune, Diana
AU - Toro, Magaly
AU - Hegde, Narasimha
AU - Meng, Jianghong
N1 - Publisher Copyright:
© 2015 by the authors.
PY - 2015
Y1 - 2015
N2 - The DNA sequence of the O-antigen gene clusters of Escherichia coli serogroups O62, O68, O131, O140, O142, and O163 was determined, and primers based on the wzx (O-antigen flippase) and/or wzy (O-antigen polymerase) genes within the O-antigen gene clusters were designed and used in PCR assays to identify each serogroup. Specificity was tested with E. coli reference strains, field isolates belonging to the target serogroups, and non-E. coli bacteria. The PCR assays were highly specific for the respective serogroups; however, the PCR assay targeting the O62 wzx gene reacted positively with strains belonging to E. coli O68, which was determined by serotyping. Analysis of the O-antigen gene cluster sequences of serogroups O62 and O68 reference strains showed that they were 94% identical at the nucleotide level, although O62 contained an insertion sequence (IS) element located between the rmlA and rmlC genes within the O-antigen gene cluster. A PCR assay targeting the rmlA and rmlC genes flanking the IS element was used to differentiate O62 and O68 serogroups. The PCR assays developed in this study can be used for the detection and identification of E. coli O62/O68, O131, O140, O142, and O163 strains isolated from different sources.
AB - The DNA sequence of the O-antigen gene clusters of Escherichia coli serogroups O62, O68, O131, O140, O142, and O163 was determined, and primers based on the wzx (O-antigen flippase) and/or wzy (O-antigen polymerase) genes within the O-antigen gene clusters were designed and used in PCR assays to identify each serogroup. Specificity was tested with E. coli reference strains, field isolates belonging to the target serogroups, and non-E. coli bacteria. The PCR assays were highly specific for the respective serogroups; however, the PCR assay targeting the O62 wzx gene reacted positively with strains belonging to E. coli O68, which was determined by serotyping. Analysis of the O-antigen gene cluster sequences of serogroups O62 and O68 reference strains showed that they were 94% identical at the nucleotide level, although O62 contained an insertion sequence (IS) element located between the rmlA and rmlC genes within the O-antigen gene cluster. A PCR assay targeting the rmlA and rmlC genes flanking the IS element was used to differentiate O62 and O68 serogroups. The PCR assays developed in this study can be used for the detection and identification of E. coli O62/O68, O131, O140, O142, and O163 strains isolated from different sources.
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U2 - 10.3390/bios5010051
DO - 10.3390/bios5010051
M3 - Article
C2 - 25664526
AN - SCOPUS:84925946914
SN - 2079-6374
VL - 5
SP - 51
EP - 68
JO - Biosensors
JF - Biosensors
IS - 1
ER -