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The diagnosis of human cases of tularemia often relies upon the demonstration of an antibody response to Francisella tularensis or the direct culturing of the bacteria from the patient. Antibody response is not detectable until 2 weeks or more after infection, and culturing requires special media and suspicion of tularemia. In addition, handling live Francisella poses a risk to laboratory personnel due to the highly infectious nature of this pathogen. In an effort to develop a rapid diagnostic assay for tularemia, we investigated the use of TaqMan 5 hydrolysis fluorogenic PCR to detect the organism in tissues of infected mice. Mice were infected to produce respiratory tularemia. The fopA and tul4 genes of F. tularensis were amplified from infected spleen, lung, liver, and kidney tissues sampled over a 5-day period. The samples were analyzed using the laboratory-based Applied Biosystems International 7900 and the Smiths Detection-Edgewood BioSeeq, a hand-held portable fluorescence thermocycler designed for use in the field. A comparison of culturing and PCR for detection of bacteria in infected tissues shows that culturing was more sensitive than PCR. However, the results for culture take 72 h, whereas PCR results were available within 4 h. PCR was able to detect infection in all the tissues tested. Lung tissue showed the earliest response at 2 days when tested with the ABI 7900 and in 3 days when tested with the BioSeeq. The results were in agreement between the ABI 7900 and the BioSeeq when presented with the same sample. Template preparation may account for the loss of sensitivity compared to culturing techniques. The hand-held BioSeeq thermocycler shows promise as an expedient means of forward diagnosis of infection in the field.

Introduction:Emergency medical services (EMS) responses to mass gatherings have been described frequently, but there are few reports describing the response to a single-day gathering of large magnitude.Objective:This report describes the EMS response to the largest single-day, ticketed concert held in North America: the 2003 “Toronto Rocks!” Rolling Stones Concert.Methods:Medical care was provided by paramedics, physicians, and nurses. Care sites included ambulances, medically equipped, all-terrain vehicles, bicycle paramedic units, first-aid tents, and a 124-bed medical facility that included a field hospital and a rehydration unit. Records from the first-aid tents, ambulances, paramedic teams, and rehydration unit were obtained. Data abstracted included patient demographics, chief complaint, time of incident, treatment, and disposition.Results:More than 450,000 people attended the concert and 1,870 sought medical care (42/10,000 attendees). No record was kept for the 665 attendees simply requesting water, sunscreen, or bandages. Of the remaining 1,205 patients, the average of the ages was 28 ±11 years, and 61% were female. Seven-hundred, ninety-five patients (66%) were cared for at one of the first-aid tents. Physicians at the tents assisted in patient management and disposition when crowds restricted ambulance movement. Common complaints included headache (321 patients; 27%), heat-related complaints (148; 12%), nausea or vomiting (91; 7.6%), musculoskeletal complaints (83; 6.9%), and breathing problems (79; 6.6%). Peak activity occurred between 14:00 and 19:00 hours, when 102 patients per hour sought medical attention. Twenty-four patients (0.5/10,000) were transferred to off-site hospitals.Conclusions:This report on the EMS response, outcomes, and role of the physicians at a large single-day mass gathering may assist EMS planners at future events.