At the same time, investigators had a fortuitous break: one of the patients had purchased two cantaloupes, one stored as cut chunks in the refrigerator and the other still uncut. The scientists prepared to test these immediately, while two staff members went grocery shopping. The pair went to three stores—one outlet of store A and two outlets of store B—returning with 15 Colorado-grown cantaloupes.
In the meantime, the patient count continued to rise. Speed was all important.
“We fast-tracked everything to the limits of the protocol,” said Maguire. “If the protocol said incubate for four to 16 hours, we incubated for four hours on the dot.”
The scientists performed PFGE analysis in parallel with MLVA, both running at max speed.
They bagged each cantaloupe with an enrichment broth and let it incubate. Then they plated the broth on agar growth media and waited for dark blue dots to appear, a tell-tale sign of Listeria.
Working over the weekend, food microbiologist Skip Gossack recovered and confirmed those Listeria colonies, while molecular biologist Justin Nucci, MPH, initiated MLVA.
The results? All five melons from store A tested negative for L. monocytogenes. All ten from store B tested positive. This information alone, said Maguire, exonerated some cantaloupe farms on the list of suspected sources.
Even more importantly, all of the distinct Listeria fingerprints associated with patient isolates were represented in the isolates from store B cantaloupes. Maguire said, “This eliminated concerns that there might be an alternate source of Listeria. Everything we were seeing in the patients, we were seeing in the cantaloupe.”
The PFGE data, coming a day or so later, was “100% concordant” with MLVA data.
As the link to cantaloupe developed, regulators were already busy with traceback investigations. Ultimately, they identified one farm as the sole source. On September 14, the farm issued a voluntary cantaloupe recall.
However, since L. monocytogenes can incubate for up to 70 days before onset of illness, cases continued to mount. By September 29, 84 cases from 19 states were reported to the CDC, among them, 15 deaths.
The good news is that the contamination did not occur three years earlier, before the CDPHE laboratory had the benefit of cooperative agreement funding from the US Food and Drug Administration and Food Emergency Response Network. Maguire said, “Without this support, the instrumentation, training and collaborative relationships [critical to breaking the investigation] would not have existed, because we wouldn’t have had the money to put them in place.”
Instead, with the infrastructure up and running, the laboratory moved quickly. Maguire said it met its major challenge: “how to identify a dangerous product but not impact the entire Colorado cantaloupe industry.”
Colorado Department of Public Health & Environment, Laboratory Division Scientific Team Responsible for Testing During the Listeria Outbreak, October 2011. (Left to right) Garth Summers (temporary-Molecular lab), Kristin Mayo (lab technician-Molecular lab), Skip Gossack (scientist-Environmental Microbiology lab), Margot Stuchin (APHL fellow-Molecular lab) and Justin Nucci (scientist-Molecular lab).