Florida Bureau of Public Health laboratories

STEC

Identification of shiga toxin-producing E. coli by microbiological and serological methods with confirmation by PCR. In order to provide pure isolates for PCR stx1 and stx2 testing, isolates are recovered from stool samples provided in Cary-Blair media or GN-Broth using PCR, pulsed-field gel electrophoresis (PFGE) and whole genome sequencing (WGS).

Analyte(s)

Shiga toxin-producing E. coli

E. coli Information

Test Code

2301 and 2302

Testing Location

Jacksonville Laboratory

Testing Department

Microbiology

Pre-approval Required?

No

Required Forms

BPHL Test Requisition, DH1847
(not needed if orders are placed by electronic order)

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Supply Ordering

Supplies may be requested from your servicing laboratory. Supplies ordered from the BPHL should only be used for specimen sent to the BPHL for testing.

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Turn Around time

5 – 7 business days after receipt at the laboratory

Minimum Volume

5 mL of raw stool, or stool sample in GN Broth or Cary Blair (preferred)

Collection Media

Sterile stool container, GN Broth, or Cary Blair

Specimen Sources

Stool

Specimen Labeling

Specimen must be labeled with at least two unique patient identifiers, e.g., name and DOB.

Information on the specimen must match the requisition and/or test order. 

Special SPecimen Preparation

This test order is for specimens predetermined to be positive for O157 and/or shiga toxin positive.

Cultures will also be run by PCR to determine detection of eae, ehx A, stx 1 and stx 2. Sero-grouping is performed for O111, O103, O26, O145, O45, O121. Serotyping performed for O157:H7.

Storage Conditions

Store specimens at 15 – 30o C until time of shipment.

Packaging and Shipping

Specimens should be shipped at 15 – 30o C.

Raw stool samples need to be received at the laboratory within 3 days of collection.

Stool specimens in Cary-Blair shipped overnight within 96 hours of collection.

Fecal matter that has been incubated at 37 °C in a GN broth for 24 hours should be shipped overnight within 24 to 48 hours of inoculation.

Include paperwork with specimens but separate them physically by placing the paperwork in the outer pocket of the biohazard bag.

unsatisfactory specimen

1) GN Broth and Cary Blair specimens received greater than 10 days from collection
2) Raw stool specimens received greater than 3 days from collection
3) No patient identification on specimen tube
4) Patient identifiers on the specimen do not match the requisition or test order
5) Specimen damaged or leaked in transit

Interferences and Limitations

Non-viable, mixed cultures may not yield reportable results

Reference Range

Negative: No pathogen detected

Result Indicator

Positive: Isolation in culture and organism ID after culture

Negative: No pathogen detected

Reference lab

CDC

Reflex testing

Whole Genome Sequencing (WGS)