Product Instructions: AlerTox Sticks Sesame

1. Intended Use

The AlerTox® Sticks Sesame Kit is a rapid, immunochromatographic, lateral flow test for the qualitative detection of sesame 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 Seeds, Sesame, and Walnut Kits, but not with other AlerTox Sticks Kits. Please read all instructions before beginning the assay.

2. Introduction

Sesame is an herbaceous plant belonging to the family Pedaliaceae, with common species being Sesamum indicum (white sesame) and Sesamum radiatum (black sesame). Allergies to sesame seeds and products can cause symptoms ranging from mild oral allergies to severe systemic reactions like anaphylactic shock. Sesame allergy prevalence is estimated at 0.1–0.9%, being more common in Asian countries. Sensitized individuals may cross-react with peanuts, walnuts, hazelnuts, rye, and poppy seeds. Separate AlerTox Sticks Kits for peanuts (KIT3094), walnuts (KIT3092), and hazelnuts (KIT3035) are available.

The European Food Safety Authority (EFSA) lists sesame seeds as an allergen requiring indication on food labels per Regulation (EU) No. 1169/2011 Annex II. Australia, Canada, Israel, and the UK also consider sesame a major food allergen. In the US, sesame allergies are the ninth most frequent, particularly affecting young children, with mandatory labeling required as of January 1, 2023.

3. Test Applications, Specificity and Sensitivity

The AlerTox Sticks Sesame Kit uses monoclonal antibodies against sesame antigen (2S seed storage protein, allergen Ses i2) to detect antigens in white and black sesame. It is suitable for:

• Food samples
• Rinse water testing
• Surface testing

For sesame seed protein, the Limit of Detection (LOD) is 3 ppm (3 mg/kg or mg/L) and the Range of Detection (ROD) is 3–220,000 ppm. Overloading (signal decrease) may occur at 1,000–220,000 ppm, but no total hook effect (false negative) is observed within this range.

For peeled sesame, the LOD is 10 ppm and the ROD is 10–250,000 ppm. Overloading may occur at 20,000–250,000 ppm, with no total hook effect observed.

On dry surfaces, the LOD is approximately 3.5 µg of sesame protein/16 cm². Specific LODs for surface testing are available on the Certificate of Analysis (search by lot number at www.hygiena.com/documents).

Section 12 lists matrices validated for the kit using an LOD of 3 ppm. AlerTox Sticks Sesame does NOT detect antigens from cereals, legumes, or other seeds. A separate kit for mustard seeds (KIT3040) is available. This is a qualitative assay; for quantification, use the AlerTox ELISA Sesame Kit (KIT3051).

NOTE:

• Sensitivity may decrease in environments rich in fats (e.g., with oil or creams).
• The kit detects proteins and peptides, not oils (e.g., sesame oil). Protein residues may remain in sesame oil. Verify results with other methods like AlerTox ELISA Sesame Kit (KIT3051) if necessary.
• Viscous, dense, or high-fat samples may affect migration along the chromatography membrane, potentially weakening or suppressing test and control lines. Contact Hygiena for information on sample extractions requiring larger dilutions that affect LOD (www.hygiena.com/support).

4. Kit Contents

5. Other Materials Not Supplied

• Grinder, mortar, or homogenization system to crush the sample
• Vortex mixer/shaker (recommended)
• Scissors (for surface sampling)
• Optional: Digital scale (sensitive to 0.1 g)

6. Precautions

• Store all kit components at 10 to 30 °C (50 to 86°F).
• Use the test strip within 10 minutes after opening the foil pouch.
• Do NOT touch the white end of the test strip.
• Do not use a test strip if it is broken, damaged, or if its pouch is torn.
• Do not use test strips beyond the expiration date.
• All test kit components are disposable; do not reuse them.
• 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 is designed to detect the target antigen in:

• Solid food
• Liquid samples (Beverages, Wash water from cutting equipment)
• Surfaces

8. Test Procedure for Solid Food Samples

1. Before adding the sample to a yellow-capped tube, mash or crush it to obtain the finest crumbs possible using a mortar or grinder.
2. Add 1 g of the sample to the yellow-capped tube. Alternatively, use the provided single-use spoons as per the chart:
   • Flours, fine powders (e.g., corn flour, rice flour, milk powder, spices): 2 spoonfuls
   • Fine crumbs (e.g., bread, cookies, cakes, snacks): 2 spoonfuls
   • Meat, fish, and cured meat (e.g., meat, fish, sausage, black pudding, pâté, canned meat and fish): 1 spoonful
3. Pour the entire contents of a blue-capped tube (10 mL) into the yellow-capped tube. Keep the blue cap for later use.
4. Close the yellow-capped tube and shake the sample for at least 20 seconds using a vortex mixer or vigorously by hand to ensure homogenization.
5. Let the mixture rest for 2 minutes for solids to settle.
6. Use a small pipette to fill the blue cap with supernatant. For samples with high fat content, avoid the fat layer of the supernatant.
7. Open the foil pouch, carefully remove the test strip by holding its blue end, and place the white end of the strip into the blue cap. Do NOT touch the white end of the test strip.
8. Wait 10 minutes to read the result. Do not read results after more than 10 minutes, as results may vary. Do not touch the test strip while waiting.

Workflow Description:

1. Crush/mash sample.
2. Add sample to yellow-capped tube.
3. Add buffer from blue-capped tube.
4. Close and shake for 20 seconds.
5. Let settle for 2 minutes.
6. Collect supernatant with a small pipette.
7. Insert test strip into the blue cap.
8. Read result after 10 minutes (Positive/Negative lines).

9. Test Procedure for Liquid Samples

Liquid samples (beverages, wash water, etc.) may be tested directly. Turbid samples should be filtered or allowed to settle.

1. Using a 3 mL pipette, add 3 mL of liquid sample to a yellow-capped tube. For thick samples (e.g., yogurt, sauce), use the provided single-use spoons (3 spoonfuls for liquid and sauces). Shake the sample to ensure homogeneity.
2. Add 3 mL of sample extraction buffer from the blue-capped tube to the sample using the 3 mL pipette. Keep the blue cap for later use.
3. Close the yellow-capped tube and shake the sample for at least 20 seconds using a vortex mixer or vigorously by hand to ensure homogenization.
4. If the liquid is cloudy, let it settle for 2 minutes.
5. Use a small pipette to fill the blue cap with supernatant. For samples with high fat content, avoid the fat layer.
6. Open the foil pouch, carefully remove the test strip by holding its blue end, and place the white end of the strip into the blue cap. Do NOT touch the white end of the test strip.
7. Wait 10 minutes to read the result. Do not read results after more than 10 minutes, as results may vary. Do not touch the test strip while waiting.

Workflow Description:

1. Add 3 mL liquid sample to yellow-capped tube.
2. Add 3 mL buffer from blue-capped tube.
3. Close and shake for 20 seconds.
4. Let settle for 2 minutes (if cloudy).
5. Collect supernatant with a small pipette.
6. Insert test strip into the blue cap.
7. Read result after 10 minutes (Positive/Negative lines).

10. Test Procedure for Surface Analysis

Collect each sample using a clean, unused swab. The swab can be used on working surfaces or equipment.

1. 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 line of 40 cm/15.6 in). For irregular surfaces, ensure swabbing technique is consistent. The area must be representative of the total area of interest. Swab an approximately 4 cm x 4 cm (1.6 in x 1.6 in) square area when possible.
2. Place the swab into the sample collection tube and trim it using scissors. Ensure the swab fits in the yellow-capped tube when the cap is closed.
3. 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 for later use.
4. Close the yellow-capped tube and shake the sample for at least 20 seconds using a vortex mixer or vigorously by hand to ensure homogenization.
5. Use a small pipette to fill the blue cap with supernatant.
6. Open the foil pouch, carefully remove the test strip by holding its blue end, and place the white end of the strip into the blue cap. Do NOT touch the white end of the test strip.
7. Wait 10 minutes to read the result. Do not read results after more than 10 minutes, as results may vary. Do not touch the test strip while waiting.

Workflow Description:

1. Wet swab in buffer.
2. Rub swab on surface.
3. Place swab in collection tube and trim.
4. Add buffer from blue-capped tube to yellow-capped tube.
5. Press swab tip against tube wall.
6. Close and shake for 20 seconds.
7. Collect supernatant with a small pipette.
8. Insert test strip into the blue cap.
9. Read result after 10 minutes (Positive/Negative lines).

11. Interpretation of Results

The test result is POSITIVE if TWO colored lines appear: one in the control zone (C) and one in the test zone (T). The color intensity of the test line may vary but is not necessarily proportional to the concentration of sesame antigen.

The test result is NEGATIVE if only ONE colored line is clearly visible in the control zone (C).

If NO colored line appears in the control zone (C), the test is INVALID.

If the test is invalid, check for and repeat the test with another strip:

• Correct specimen handling
• Correct test procedure
• Expiration date
• Correct storage conditions

For further assistance, contact Hygiena at www.hygiena.com/support.

IMPORTANT NOTE! AlerTox Sticks is a qualitative test for screening samples for internal quality control. It cannot replace laboratory analysis for quantification.

12. Validation

AlerTox Sticks Sesame has been validated for the following matrices:

Matrices should be validated before use. For additional information about matrix validation, contact Hygiena at www.hygiena.com/support.

13. Disclaimer

Field of use: Use the Hygiena product 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 should not be the sole basis for assessing product safety for release to consumers. Data must not be used for human diagnostic or treatment purposes. Read the Limitation of Warranty and Liability (available in Hygiena General Terms and Conditions at www.hygiena.com/terms-and-conditions) before use.

Products are made from high-quality raw materials. No warranty is made regarding suitability other than to measure target antigen content when used exactly per instructions, except regarding material quality.

Use of the kit for any other purpose is outside its intended use. For unvalidated matrices, Hygiena cannot guarantee fitness for purpose or accuracy of results. Users choosing to use the product on unvalidated matrices should perform their own fit-for-use testing. Hygiena's liability for damages is limited to the replacement value of the kit.

For additional information or assistance with matrix validation, contact Hygiena at www.hygiena.com/support. 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

• INS3031 REVA, October 2021: The initial protocol.
• INS-KIT3096-001-REVA, July 2025: Updated the LOD and ROD information. Standardized branding, wording, some graphic workflows, and document ID number.