Detection of Shigella in feces using DNA amplification.
A rapid diagnostic method employing a polymerase chain reaction procedure (PCR) was used to identify Shigella and enteroinvasive Escherichia coli. This procedure amplified a region of the invasive-associated locus (ial) from a crude DNA extract of feces. A synthetic 21-base oligonucleotide corresponding to the ial gene sequence was shown to specifically hybridize only with enteroinvasive E. coli (EIEC) strains and Shigella species. Upon PCR amplification, a 320-base pair fragment was generated in DNA extracted from feces reconstituted with EIEC or Shigella flexneri but not in DNA from 70 normal stools lacking these organisms and could be readily detected by the ial probe. For identifying Shigella and EIEC, the PCR assay was 10(5)- and 10(2)-fold more sensitive than standard biochemical tests and the macrocolony hybridization assay, respectively. These findings demonstrate a novel methodology for rapid, sensitive, and culture-independent diagnosis of diarrhea caused by these pathogens and underscores the utility of PCR in the diagnostic laboratory.
Department of Microbiology, Stanford University, CA 94305.
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G
Frankel
L
L
Riley
J A
JA
Giron
J
J
Valmassoi
A
A
Friedmann
N
N
Strockbine
S
S
Falkow
G K
GK
Schoolnik
D001483:Base Sequence; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D003967:Diarrhea / Q000175:diagnosis*; D004405:Dysentery, Bacillary / Q000175:diagnosis*; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification; D004927:Escherichia coli Infections / Q000175:diagnosis; D005243:Feces / Q000382:microbiology*; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D012760:Shigella / Q000235:genetics / Q000302:isolation & purification*; D012763:Shigella flexneri / Q000235:genetics / Q000302:isolation & purification
D004269:DNA, Bacterial; D015345:Oligonucleotide Probes