Chemistry: molecular biology and microbiology – Micro-organism – per se ; compositions thereof; proces of... – Bacteria or actinomycetales; media therefor
Patent
1997-03-26
1998-08-25
Wax, Robert A.
Chemistry: molecular biology and microbiology
Micro-organism, per se ; compositions thereof; proces of...
Bacteria or actinomycetales; media therefor
4353201, 536 237, C12N 121, C12N 1563, C07H 2104
Patent
active
057982600
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The invention relates to genetic engineering and particularly to the demonstration that a contiguous segment of chromosomal DNA from E. coli O157:H7 encodes an adhesin that mediates colonization of the gastrointestinal tracts of bovines, and possibly humans, with E. coli O157:H7 and bacteria using structurally related adherence mechanisms.
BACKGROUND OF THE INVENTION
E. coli O157:H7 is a virulent and common foodborne pathogen. Most outbreaks, and many sporadic cases (38,42; see the appended Citations), have been attributed to food of bovine origin. Most E. coli O157:H7 infections are sporadic, but this organism can cause massive epidemics by contamination of ground beef (19) and water (69). E. coli O157:H7 is transmissible from person to person, but the disappearance of the strain which caused the massive 1993 outbreak in Washington State soon after recall of the incriminated vehicle demonstrates that ingestion of contaminated beef, and not person to person spread, is the chief source of human infection.
E. coli O157:H7 organism elaborates Shiga-like toxins (SLT) I and/or II. SLT I and II inhibit protein synthesis by disrupting a glycosidic bond at a specific adenine (A4324) in 28S rRNA of the 60S ribosomal subunit. SLT-producing E. coli (SLTEC) are ubiquitous in food (62) and animals (47). The vast majority are probably not human pathogens.
Current data suggest that E. coli O157:H7 is the most common and medically significant SLTEC. Only one outbreak of bloody diarrhea caused by SLTEC other than E. coli O157:H7 has ever been reported (11). Additionally, even when sought appropriately, non-O157:H7 SLTEC are rarely found in stools submitted for bacterial culture in North America compared to their frequency in the environment (8,52,59). Moreover, E. coli O157:H7 is the predominant precipitant of the hemolytic uremic syndrome (HUS), the most important complication of enteric infection with E. coli O157:H7. For example, E. coli O157:H7 was found in 96% of HUS patients if stool was obtained within the first six days of diarrhea (72). Even though non-O157:H7 SLTEC have caused some cases of HUS in several foreign series (10,11,12,35,40), these strains have never been reported to cause HUS in the United States. These data suggest that E. coli O157:H7 is the most important cattle-borne human pathogen threatening the food supply of this country today.
Cattle are the only reservoir of E. coli O157:H7 so far identified. Approximately 1 in200 apparently healthy northwestern United States dairy and beef cattle carry E. coli O157:H7, and 8 to 16% of herds have at least one infected animal (25). Similar carriage rates have been detected nationwide (26). These are probably minimum carriage rates, because the technique used to culture E. coli O157:H7 is relatively insensitive.
A very low inoculum of E. coli O157:H7 can cause human disease. Person to person spread occurs rather easily in outbreaks and among sporadic cases (5,6,60). Microbiologic analysis of the contaminated hamburger from the 1993 Western United States outbreak demonstrated that only approximately 200 E. coli O157:H7 were present in each of the contaminated patties (46). It is probable that the inadequate cooking that was applied reduced this concentration by at least one log, suggesting that very few E. coli O157:H7, perhaps in the range of 1-10 bacteria, can cause clinically apparent infection.
Data suggest that the incidence of diseases caused by E. coli O157:H7 has increased in the United States, independent of ascertainment bias by diagnosing physicians (44,70). Additionally, an increasing rate of antibiotic resistance in Washington State human isolates of E. coli O157:H7 might portend an increased prevalence of this pathogen in animals administered antibiotics. For example, before 1988, none of 56 strains of E. coli O157:H7 were resistant to a wide variety of antibiotics tested, whereas after 1988, 7.4% of 176 strains were resistant to the same combination of antimicrobials (streptomycin, sulfamethoxazole, and tetracycline).
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Besser Thomas E.
Bilge Sima S.
Tarr Phillip I.
Vary, Jr. James C.
Children's Hospital and Medical Center
Lau Kowai
University of Washington
Washington State University Research Foundation
Wax Robert A.
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