Multiplex polymerase chain reaction method discriminating Escherichia coli and Shigella sp.
To distinguish between Escherichia coli and other bacteria that have similar biochemical characteristics, 3 polymerase chain reaction techniques were combined. The primer sets cydA-F2-A2 and cydA-R2-A2 were designed to amplify 605 base pairs of nucleotide sequence speciﬁc for the cydA gene of Escherichia coli; primer sets lacZ-F-A and lacZ-R-A to amplify 1,023 bp of nucleotide sequence speciﬁc for the lacZ gene of Escherichia coli; and primers lacA-F2-A2 and lacA-R2-A2 to amplify 325 bp of nucleotide sequence speciﬁc for the lacA gene of Escherichia coli. As a result, 3 nucleotide fragments were generated when 3 samples DNA from Escherichia coli were used as template. On the other hand, 1,023- and 605-bp products were obtained when DNA of Shigella sonnei was used, and a 605-bp product was obtained when DNA of Shigella ﬂexneri was used. The speciﬁcity of the technique was conﬁrmed by comparing it with the conventional culture test; the consistency rate of both tests was 0.749. These results suggest that the technique described in the present study will be useful for distinguishing Escherichia coli from Shigella species with accuracy and speciﬁcity.
Ibaraki prefectural institute of public health, 993-2, Kasarhara-cho, Mito, Ibaraki 310-0852, Japan. y.yamazaki@pref.ibaraki.lg.jp
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Yamazaki
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Fukasawa
D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012760:Shigella / Q000235:genetics / Q000302:isolation & purification*
D017931:DNA Primers