AlerTox Sticks Crustacean
Product Instructions
Brand: Hygiena
Reference: KIT3036
1. Intended Use
The AlerTox® Sticks Crustacean Kit is a rapid, immunochromatographic, lateral flow test for the qualitative detection of crustacean antigen in food, kitchens, and production facilities. Samples prepared according to these instructions can be tested using test strips from AlerTox Sticks Almond, Crustacean, Fish, Hazelnut, Mustard Seed, Sesame, and Walnut Kits, but not with other AlerTox Sticks kits. Read all instructions before beginning the assay.
2. Introduction
Crustaceans (Crustacea) are a taxonomic division (subphylum) of arthropods, including insects and spiders. They are common in seas and fresh waters, with many species also living on land. Crustaceans are a significant part of seafood consumption globally, and crustacean allergies can cause symptoms ranging from mild oral reactions to severe, life-threatening systemic reactions like anaphylactic shock or bronchial asthma. Allergy to crustaceans is among the top 10 food allergies in some regions. In the US, the Food Allergen Labeling and Consumer Protection Act (FALCPA) identifies crustacean allergy as a major food allergy, requiring specific labeling. The European Food Safety Authority (EFSA) lists crustaceans as allergens requiring indication on food products. Detection of crustacean protein can also support religious food-labeling requirements.
3. Test Applications, Specificity and Sensitivity
The AlerTox Sticks Crustacean Kit uses monoclonal antibodies against tropomyosin, a major antigen in crustacean muscle, identified as allergen Met e 1 (shrimp) and Cra c 1 (North Sea shrimp), and similar proteins in other species. This kit is suitable for:
- Food samples
- Rinse water testing
- Surface testing
The Limit of Detection (LOD) is 10 ppm of dried raw shrimp protein (10 mg/kg or mg/L). The Range of Detection (ROD) is 10 – 100,000 ppm. Overloading (signal decrease) may occur at 5,000 – 100,000 ppm. A hook effect (false negative) may appear above 250,000 ppm; repeat the test with a diluted sample if suspected.
View LODs for surface testing on the Certificate of Analysis at www.hygiena.com/documents. Section 12 lists validated matrices for the kit at 10 ppm LOD.
AlerTox Sticks Crustacean does NOT detect antigens from fish, mollusks, or other meats. Separate kits are available for fish (KIT3038). For quantifying antigen amounts, use the AlerTox ELISA Crustacean Kit (KIT3059).
Note:
- AlerTox Sticks Crustacean can detect antigens of phylum Arthropoda (insects, mites, spiders). Sensitivity to these cross-reactive antigens is lower (100-1,000 times) than for shrimp. It can detect cross-contamination by insects in foods like grains or flour.
- Sensitivity decreases in environments rich in fats (e.g., oil or creams).
- The kit detects proteins and peptides, not oils (e.g., krill oil). Verify results with other methods for krill oil if protein residues are suspected.
- Viscous, dense, or high-fat samples may affect migration and assay results. Contact Hygiena support for information on sample extraction and dilutions.
4. Kit Contents
| Component | KIT3036 |
|---|---|
| Crustacean immunochromatographic test strips individually packaged in foil pouches | 10 |
| Sample collection tubes with yellow caps | 10 |
| Sample extraction buffer in tubes with blue caps, 10 mL | 10 |
| Spoons | 10 |
| Small pipettes | 10 |
| Pipettes, 3 mL (only for testing liquid samples) | 10 |
| Swabs (only for testing surfaces) | 10 |
5. Other Materials Not Supplied
- AlerTox Polyphenol Additive (Product No. ASY3213) for samples with polyphenols/antioxidants (e.g., 70% chocolate, coffee).
- Grinder, mortar, or homogenization system for crushing samples.
- Vortex mixer/shaker (recommended).
- Scissors (for surface sampling).
- Digital scale (sensitive to 0.1 g) (optional).
6. Precautions
- Store kit components at 10 to 30 °C (50 to 86°F).
- Use test strip within 10 minutes after opening the foil pouch.
- Do NOT touch the white end of the test strip.
- Do not use a broken, damaged, or improperly packaged test strip.
- Do not use test strips beyond the expiration date.
- All kit components are disposable; do not reuse.
- Do not combine components from different kits.
7. Sample Handling
All samples must be at 18 to 35 °C (64.4 to 95 °F) before use. The test detects the target antigen in:
- Solid food
- Liquid samples (Beverages, Wash water from cutting equipment)
- Surfaces
8. Test Procedure for Solid Food Samples
Important: If the sample contains polyphenols or antioxidants (e.g., 70% chocolate, coffee), add 1 spoonful of AlerTox Polyphenol Additive (ASY3213) to a yellow-capped tube before proceeding.
Step 8.1: Sample Preparation
Mash or crush the sample to obtain the finest crumbs possible using a mortar or grinder.
Step 8.2: Add Sample
Add 1 g of the sample to the yellow-capped tube. Alternatively, use the following chart for equivalent amounts:
| Food Type | Examples | Spoonfuls |
|---|---|---|
| Flours, fine powders | Corn flour, rice flour, milk powder, spices | 2 |
| Fine crumbs | Bread, cookies, cakes, snacks | 2 |
| Meat, fish and cured meat | Meat, fish, sausage, black pudding, pâté, canned meat and fish | 1 |
Step 8.3: Add Buffer
Pour the entire contents of a blue-capped tube (10 mL) into the yellow-capped tube. Keep the blue cap for later use.
Step 8.4: Homogenize
Close the yellow-capped tube and shake for at least 20 seconds using a vortex mixer or vigorously by hand for homogenization.
Step 8.5: Settle
Let the sample rest for 2 minutes for solids to settle.
Step 8.6: Collect Supernatant
Use a small pipette to fill the blue cap with supernatant. For high-fat samples, avoid the fat layer.
Step 8.7: Insert Test Strip
Open the foil pouch, remove the test strip by its BLUE end, and place the white end into the blue cap. Do NOT touch the white end.
Step 8.8: Read Result
Wait 10 minutes to read the result. Do not read after more than 10 minutes, as results may vary. Do not touch the test strip while waiting.
9. Test Procedure for Liquid Samples
Liquid samples (beverages, wash water, etc.) can be tested directly. Turbid samples should be filtered or allowed to settle.
Important: If the sample contains polyphenols or antioxidants (e.g., 70% chocolate, coffee), add 1 spoonful of AlerTox Polyphenol Additive (ASY3213) to a yellow-capped tube before proceeding.
Step 9.1: Add Sample
Using a 3 mL pipette, add 3 mL of liquid sample to a yellow-capped tube. For thick samples (yogurt, sauce), use the chart for equivalent amounts:
| Food Type | Examples | Spoonfuls |
|---|---|---|
| Liquid and sauces | Milk, juice, condensed milk, yogurt, soup, gravy, sauce, cream | 3 |
Shake the sample to ensure homogeneity.
Step 9.2: Add Buffer
Add 3 mL of sample extraction buffer from the blue-capped tube to the sample using the 3 mL pipette. Keep the blue cap.
Step 9.3: Homogenize
Close the yellow-capped tube and shake for at least 20 seconds using a vortex mixer or vigorously by hand.
Step 9.4: Settle (if cloudy)
If the liquid is cloudy, let it settle for 2 minutes.
Step 9.5: Collect Supernatant
Use a small pipette to fill the blue cap with supernatant. For high-fat samples, avoid the fat layer.
Step 9.6: Insert Test Strip
Open the foil pouch, remove the test strip by its BLUE end, and place the white end into the blue cap. Do NOT touch the white end.
Step 9.7: Read Result
Wait 10 minutes to read the result. Do not read after more than 10 minutes, as results may vary. Do not touch the test strip while waiting.
10. Test Procedure for Surface Analysis
Collect samples using a clean, unused swab on working surfaces or equipment.
Step 10.1: Swab Surface
Wet the swab by dipping it in the blue-capped tube. Firmly rub and rotate the swab on the testing surface using a zigzag pattern (at least 16 cm² / 2.5 in² or a 40 cm / 15.6 in line). For irregular surfaces, maintain consistent swabbing. The area must be representative of the total area of interest.
Step 10.2: Prepare Swab
Place the swab into the sample collection tube and trim it using scissors. Ensure the swab fits in the yellow-capped tube when closed.
Step 10.3: Add Buffer and Extract
Pour the entire contents of a blue-capped tube (10 mL) into the yellow-capped tube. Press the swab tip against the inside wall of the yellow-capped tube to facilitate sample extraction into the buffer. Keep the blue cap.
Step 10.4: Homogenize
Close the yellow-capped tube and shake for at least 20 seconds using a vortex mixer or vigorously by hand.
Step 10.5: Collect Supernatant
Use a small pipette to fill the blue cap with supernatant.
Step 10.6: Insert Test Strip
Open the foil pouch, remove the test strip by its BLUE end, and place the white end into the blue cap. Do NOT touch the white end.
Step 10.7: Read Result
Wait 10 minutes to read the result. Do not read after more than 10 minutes, as results may vary. Do not touch the test strip while waiting.
11. Interpretation of Results
POSITIVE: TWO colored lines appear: one in the control zone (C) and one in the test zone (T). The intensity of the test line may vary but is not proportional to antigen concentration.
NEGATIVE: Only ONE colored line is clearly visible in the control zone (C).
INVALID: If NO colored line appears in the control zone (C), the test is invalid. Check specimen handling, test procedure, expiration date, and storage conditions. Repeat with a new strip. For assistance, contact Hygiena at www.hygiena.com/support.
IMPORTANT NOTE: AlerTox Sticks is a qualitative test for internal quality control. It cannot replace laboratory analysis for quantification.
12. Validation
AlerTox Sticks Crustacean has been validated for the following matrices:
| Validated Matrices |
|---|
| Chocolate (70%)* |
| Coffee* |
| Fish (including canned sardines) |
| Flours |
| Dairy |
| Meat |
| Prepared food |
| Snacks |
* 70% chocolate and coffee samples were prepared using the AlerTox Polyphenol Additive.
Matrices should be validated before use. For more information on matrix validation, contact Hygiena at www.hygiena.com/support.
13. Disclaimer
Field of use: This Hygiena product is for research and development, quality assurance, and quality control under the supervision of technically qualified persons. Information generated is for use with the user's regular quality assurance program and not as the sole basis for product safety assessment for consumers. Data must not be used for human diagnostic or treatment purposes. Read the Limitation of Warranty and Liability (available at www.hygiena.com/terms-and-conditions) before use.
Products are made from high-quality raw materials. No warranty is made regarding suitability beyond measuring the target antigen content as per instructions, except for material quality.
Use for any other purpose is outside the intended use. For unvalidated matrices, Hygiena cannot guarantee fitness for purpose or accuracy of results. Users choosing unvalidated matrices should perform their own fit-for-use testing. Damages are limited to the replacement value of the kit.
For additional information or assistance with matrix validation, contact Hygiena at www.hygiena.com/support. All Hygiena Terms and Conditions apply and can be found at: www.hygiena.com/terms-and-conditions.
14. Contact Information
For more information, visit www.hygiena.com/contact. For technical support, visit www.hygiena.com/support.
15. Change Index
- INS3019 REVA, February 2019: The initial protocol.
- INS-KIT3036-001-REVA, July 2025: Updated ROD and LOD information. Included use of AlerTox Polyphenol Additive for sample preparations. Standardized branding, wording, graphic workflows, and document ID number.
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