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Shigatoxin EIA (E. coli)   

 
IMPORTANT! Pre-approval needed from state or local health department. Local or State Health Department. Send approval to NPHL@unmc.edu or call (402) 559-9444.

NPHL has moved to an electronic ordering system, NUlirt, and it is now the preferred method to order all tests. Once ordered, print a "batch list" (see NUlirt Guide).

Forms Required - Nulirt batch list or NPHL Test Request Form (completed in entirety) must accompany the specimen to include the following information: symptoms and date of onset, pregnancy status, travel and vaccine history. Testing will not be performed without all required information.

Please call NPHL Client Services at (402) 559-2440 or toll free at (800) 334-0459 for further clarification.

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Test: Shigatoxin EIA (E. coli) 
Order in NUlirt or complete NPHL Test Order Form  
Synonym: Shigatoxin testing, E. coli 0157H7, Shiga-like toxin EIA, Non-E.coli O157)  
Method: Enzyme-linked immunosorbent assay  
Availability: Test performed daily; Turnaround time 3d.  
Specimen: Stool  specimen or actively growing culture  
Collection Device: Enteric Plus Para-Pak or Cary Blair Media
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 or Cary Blair Media to bring the liquid level to the 'Fill' line or approximately 5-10mL of stool sample. Tighten the cap and shake firmly to insure that the specimen is mixed. 
Shigatoxin positive stool in pure culture of E.coli sorbitol-negative isolate on MacConkey or MacConkey with sorbitol plate for identification.

Specimen Labeling: Test subject to CLIA regulations and required 2 patient identifiers on specimen container and requisition forms  
Volume: 10mL in transport media  
Storage/Transport:

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 specimen 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 filled with frozen gel-packs to keep specimen refrigerated or dry ice to keep specimen frozen. 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.  

Ship pure isolates that are shigatoxin positive as UN 2814 Infectious Substances, affecting humans (Category A) and by FedEx. Place isolates 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 A shipping container. Position NUlirt packing list or NPHL Test Order form between secondary container and box lid. Seal with tape. Attach airway bill and Shippers Declaration to top of box. Outer box must be labeled “UN 2814 Infectious Substances, affecting humans.”   Follow appropriate DOT/IATA Regulation Guidelines.  For assistance, call NPHL State Training Coordinator at (402) 559-3590. Omaha & Lincoln laboratories can ship via NPHL ground courier by calling client services (866) 290-1406 and request exclusive Category A courier.
See instructions and shipping address: http://nphl.org/

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

 
Unacceptable: Stool received during a colonoscopy. Unpreserved stool will be rejected if greater than 1 hour 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.  
Specimen Stability: Ambient 15-25°C in transport media for 96h (2d) refrigerated or 4d.  
Reference Interval: Shigatoxin negative by EIA  
Reportable Disease: Report positive Shigatoxin results within 7d to local or state health department. Refer to Nebraska DHHS Title 173, Communicable Diseases. All shigatoxin producing E.coli isolates should be submitted to the NPHL for molecular sequencing to supplement and epidemiological investigation.  
Comments: Pre-approval needed from state or local health department.  Send approval to NPHL@unmc.edu or call (402) 559-9444.

This test will identify the presence of Shiga-like toxin E. coli in stool; however, it cannot determine the specific strain of E. coli (i.e. O157:H7).

This test can be performed either on a stool sample or on actively growing isolate of E. coli in pure culture.  

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 24h apart that have been collected no sooner than 48h following discontinuation of antibiotics or 24h after cessation of diarrhea symptoms.

NUlirt packing list or NPHL Test Order Form (completed in entirety) must accompany the specimen.  
Revised: 6/21/2021 
If you have questions about proper specimen collection, please call NPHL Client Services at (402) 559-2440 or toll free at (800) 334-0459.

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