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Introduction to Epidemiology: The Epidemiologic Approach

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Introduction to Epidemiology: The Epidemiologic Approach

Escherichia coli O157:H7 was first identified as a human pathogen in 1982 in the United States of America, following an outbreak of bloody diarrhea associated with contaminated hamburger meat. Sporadic infections and outbreaks have since been reported from many parts of the world,including North America, Western Europe, Australia, Asia, and Africa. Although other animals are capable of carrying and transmitting theinfection, cattle are the primary reservoir for E. coli O157:H7. Implicated foods are typically those derived from cattle (e.g., beef,hamburger, raw milk); however, the infection has also been transmitted through contact with infected persons, contaminated water, and other contaminated food products.

Infection with E. coli O157:H7 is diagnosed by detecting the bacterium in the stool. Most laboratories that culture stool do not routinely test for E. coli O157:H7, but require a special request from the health care provider. Only recently has E. coli O157:H7 infection become nationally notifiable in the U.S. Outside the U.S., reporting is limited to a few but increasing number of countries.

In the last week of June 1997, a certain state department of community health in the US noticed an increase in laboratory reports of E. coli O157:H7 infection. Fifty-two infections had been reported that month, compared with 18 in June of 1996. In preliminary investigations, no obvious epidemiologic linkages between the patients were found. The increase in cases continued into July.

Laboratory subtyping can help determine if an increased number of isolates of the same bacterial species results from a common source outbreak. Subtyping methods are based on selected biologic and/or geneticcharacteristics of bacteria that tend to differ between isolates of the same species. In a common source outbreak, however, isolates typically arise from the same parent organism. These isolates will be similar to each other with respect to these biologic and genetic characteristics and have similar subtyping results.

One subtyping method is DNA “fingerprinting” by Pulsed Field Gel Electrophoresis (PFGE). In DNA fingerprinting, the bacterial DNA is cut into pieces. The pieces are separated by placing them in a jelly-like substance (i.e., the gel), acting as a sieve, to which a pulsing electric fieldis applied. The electric field drives the DNA pieces across the gel over a period of hours. The smaller pieces move through the gel more quickly and the larger pieces more slowly resulting in a separation of the DNA into distinct bands. The bands are made to fluoresce and are read under ultraviolet illumination. This DNA“fingerprint” resembles a bar code. (Figure 1)

Figure 1. Typical DNA banding pattern resulting from PFGE.

Different DNA composition will result in different PFGE banding patterns. Bacteria descended from the same original parent will have virtually identical DNA and their DNA fingerprints will be in distinguishable. Identification of a cluster of isolates with the same PFGE pattern suggests that they arose from the same parent and could be from the same source.

Similar DNA fingerprints alone, however, are insufficient to establish a linkage between isolates and a common source outbreak. An epidemiologic investigation is necessary to demonstrate that there is a common source and to identify it. To be most useful, PFGE subtyping needs to be performed on a routine basis, in realtime, so that results are available (and reviewed) soon after a case is first detected.

Figure2. PFGE results on E. coli O157:H7 isolates, June-July 1997.

Typically, a PFGE “pattern” is defined as having the same banding pattern but including up to one band difference. By this definition, isolates #2, 3, 4, 6, and 7 are indistinguishable by these PFGE results. (Isolate #4 differs by one band.)

DNA finger printing, performed during the second week of July showed that 17 of the first 19 E. coli O157:H7 isolates from June-July were indistinguishable. They did not match any fingerprints from a convenience sample of isolates frompatients with E. coli O157:H7 infection before May.

Based on the PFGE findings, it was suspected the cases of E. coli O157:H7 infection resulted from a common source. On July 15, an investigation was initiated.


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