J Korean Soc Microbiol.  1998 Feb;33(1):99-110.

Use of Polymerase Chain Reaction and Serum Antibodies for Diagnosis of Enterohemorrhagic Escherichia coli

• Oh YH
• Kim YB

Abstract

Fecal isolates of Escherichia coli which were collected from diarrheal patients and HUS patient in Pusan National University Hospital between 1990 and 1996, were serotyped and analyzed for plasmid DNA profile, biotype, HEp-2 cell adherence ability, reactivity to eae probe and for production of verotoxins (VT). In order to ease the diagnosis of EHEC infection, a LPS- based solid phase enzyme linked immunosorbent assay was utilized to detect serological diagnosis of EHEC infection. The following results were obtained. Among 150 EPEC isolates and HUS patient's stool, 7 EHEC were found. The 7 EHEC belonged to 5 different serotypes 0157:H7, 0143:H-, 0166:H-, 0128:H2, 026:H-, and 0111:H 21 previously associated with human haemorrhagic colitis (HC) and haemolytic uraemic syndrome (HUS). Biochemical cheracteristic analysis indicated 7 strains were biotype 1 and was found to have siderophilins suggesting their advantagous growth in vivo. For plasmid profiles, all strains had 60 MDa plasmid and several smaller sizes of plasmids. Three strains of Escherichia coli serotype 0157:H7, 0128:H2, and 026:H- showed one pattern of adherence in the HEp-2 cell assay namely, localized adherence and were positive for eae probe when tested by colony blot hybridization assay. PCR using specific primers for VT1, VT2 was tested, and all 7 strains carried VT1 gene only. PCR products of 130-bp (VT1) and 346-bp (VT2) were successfully amplified simultaneously in a single reaction. The multiplex PCR method can be used to specifically identify EHEC. The serum obtained from HUS patient of enterohemorrhagic E. coli were analyzed for rises in titer of intibody to somatic 02, 026, 0111, 0128, 0143, 0145, 0157, and 0165. Although response to the somatic 0 correlated significantly with response to the 026 rises of antibody titer to somatic 0 in acute stage of disease and anti-VT had not so many relation to that of VT. These results suggest that ELISA can be used to detect somatic 0 in serum and it is a useful method to diagnose the infection caused by EHEC rapidly.

MeSH Terms

Antibodies*
Busan
Colitis
Diagnosis*
DNA
Enterohemorrhagic Escherichia coli*
Enteropathogenic Escherichia coli
Enzyme-Linked Immunosorbent Assay
Escherichia coli
Humans
Multiplex Polymerase Chain Reaction
Plasmids
Polymerase Chain Reaction*
Shiga Toxins
Transferrin
Antibodies
DNA
Shiga Toxins
Transferrin