Shigatoxin testing, E. coli O157H7, Shiga-like toxin EIA, Non-E.coli O157

Qualitative Enzyme Immunoassay

Test performed daily; Turnaround time 1d.

Stool specimen or actively growing isolate.

Collect stool in a clean dry container or NPHL Stool collection kit (available at Client Services). IMMEDIATELY transfer sufficient stool to Para-Pak Enteric Plus Transport vial to bring the liquid level to the 'Fill' line or approximately 10mL of stool sample. Tighten the cap and shake firmly to ensure that the specimen is mixed. 
Specimen Labeling: Test subject to CLIA regulations and requires 2 patient identifiers and specimen source on container.

10mL in transport media

Prior to and during shipment:  Ambient 15-25°C
Shipping instructions, including specimen-handling requirements during transport:  Seal screw-top tubes with Parafilm™ or tape and place in biohazard bag with absorbent material.
Place stool specimens in maintenance media inside leak proof secondary container (biohazard bag and if transporting by air, use Tyvek™ Envelope) with adsorbent material. The secondary container must be packed inside an approved Category B shipping container.  Position NUlirt packing list or NPHL Test Order form between secondary container and box lid. Seal with tape.  Label outer box as UN3373 Category B, Biological Substance. Transport by NPHL ground courier. If courier not available, ship by FedEx, UPS, or USPS (1st Class, Priority or Express Mail). Follow appropriate DOT/IATA Regulation Guidelines.  For assistance, call NPHL State Training Coordinator at (402) 559-3590.
See instructions and shipping address: http://nphl.org/

NUlirt packing list or appropriate paperwork (completed in entirety) must accompany specimens

Stool received during a colonoscopy. Unpreserved stool will be rejected if greater than 1h old. Enteric Plus inoculated with stool greater than 96hr or greater than 4d. Frozen stool. Stool contaminated with urine, barium, bismuth, mineral oil, water, or other contaminants are unacceptable. Extremely 'hard' formed stools are unacceptable. Do not use the vial if beyond expiration date or contents of the vial are cloudy or yellow.

Ambient or 2-8°C for 96hr or 4d at room temperature in Enteric Plus Vial.

Shigatoxin negative by EIA. 

Report positive Shigatoxin E.coli results within 7 days to local or state health department. Refer to Nebraska DHHS Title 173, Communicable Diseases. Ship all Shigatoxin positive E.coli isolates to NPHL.

Pre-approval needed from state or local health department.  Send approval to NPHL@unmc.edu or call (402) 559-9444.
All shigatoxin positive stools ( in enteric transport media or plate isolate) should be sent to the NPHL for isolation of the shigatoxin positive E. coli for whole genome sequencing.  Stool specimens can be sent as a Category B, biological substance. All shigatoxin positive isolates must be shipped as Category A, Infectious Substance. Order as E. coli (Hemorrhagic) Culture, Stool, HECCU.
This test will identify the presence of Shiga-like toxin E. coli in stool; however, it cannot determine the specific strain of E. coli (ie. O157:H7).
The Shiga-like toxin EIA can be used as a test for proof-of-cure testing following a primary diagnosis of enterohemorrhagic E. coli-caused infection.  Frequency for shigatoxin proof-of-cure testing can vary according to jurisdictional recommendations.  In general, cure is defined as 2 consecutive negative stools taken 24 hours apart that have been collected no sooner than 48 hours following discontinuation of antibiotics or 24 hours after cessation of diarrhea symptoms.

NUlirt packing list or NPHL Test Order Form (completed in entirety) must accompany the specimen