[Detection of two Shiga-like toxins from Escherichia coli O157:H7 isolates by the polymerase chain reaction method].
Fourteen isolates of E. coli O157:H7 and five isolates of S. dysenteriae type-1 were examined by polymerase chain reaction (PCR) for the structural genes (slt-I or slt-II), encoding Shiga-like toxins (SLTs). The two primer pairs (V1; 5'AGTTAATGTGGTGGCGAA and V2; 5'GACTGCGTCAGTGAGGTT for SLT-I, V3; 5'TTCGGTATCCTATTCCCG and V4; 5'TCTCTGGTCATTGTATTA for SLT-II) used were of the same positions representing the DNA sequence covering 471bp of the slt-I or slt-II. A 5-microliter portion of boiled bacterial culture broth was used as template DNA in a PCR-reaction mixture of 50 microliters. Two classes, slt-I alone or both slt-I and slt-II, were recognized in E. coli strains. All of S. dysenteriae type-1 strains examined contained slt-I alone. Our results indicate that PCR using these primer pairs is a simple, rapid, sensitive, and specific method and suitable for use in routine diagnostic microbiology laboratories.
Kobayashi
K
K
Osaka Prefectural Institute of Public Health, Division of Public Health.
Seto
K
K
Makino
M
M
D001427:Bacterial Toxins / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D004926:Escherichia coli; D006801:Humans; D016133:Polymerase Chain Reaction; D012762:Shigella dysenteriae
D001427:Bacterial Toxins