Rapid diagnostic method for the detection of diarrheagenic Escherichia coli by multiplex PCR.
Two novel multiplex polymerase chain reaction (mPCR) assays were originally developed for detecting nine targeted virulence-associated genes of five categorized diarrheagenic Escherichia coli (DEC). The mPCR assay 1 included five primer sets (stx1, eaeA, invE, STp gene, and astA), and assay 2 included four primer sets (stx2, aggR, STh gene, and LT gene). The two mPCRs showed 100% specificity in identifying the reference strains without nonspecific bands, and 51 DEC and 38 astA gene-positive E. coli strains from 683 E. coli-like isolates. Our mPCR methods showed high sensitivity and specificity for detecting nine virulence genes of DEC strains. We proved that these methods will contribute to reducing the cost for the reagents of mPCR reported elsewhere and could, therefore, contribute to the diagnosis of DEC in clinical laboratories.
Fujioka
Miyuki
M
Division of Medical Life Sciences, Hirosaki University Graduate School of Health Sciences, Aomori 036-8564, Japan. mfujioka@cc.hirosaki-u.ac.jp
Kasai
Kosuke
K
Miura
Tomisato
T
Sato
Tatsusuke
T
Otomo
Yoshimitsu
Y
D017931:DNA Primers / Q000235:genetics; D003967:Diarrhea / Q000382:microbiology*; D004926:Escherichia coli / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D004927:Escherichia coli Infections / Q000175:diagnosis* / Q000382:microbiology; D029968:Escherichia coli Proteins / Q000235:genetics; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D037521:Virulence Factors / Q000235:genetics
D017931:DNA Primers; D029968:Escherichia coli Proteins; D037521:Virulence Factors