Attogene AU2024-02 Microcystin Lateral Flow Kit User Manual

: June 9, 2024
: Attogene

Table of Contents

1. Intended Use
2. Introduction
3. Kit Contents
4. Storage and Stability
5. Required Materials Not Supplied
6. Precautions
7. Water Collection and Storage
8. Procedure
9. Interpretation of Results
10. Additional Analysis
References
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Attogene AU2024-02 Microcystin Lateral Flow Kit User Manual

Attogene AU2024-02 Microcystin Lateral Flow Kit User
Manual

1. Intended Use

For the rapid screening of microcyst ins in drinking water samples at or above 0.1 ppb. Samples requiring regulatory action should be confirmed by ELISA, HPLC, or other conventional methods. To protect consumers from adverse health effects caused by these toxins, the World Health Organization (WHO) has proposed a provisional upper limit for Microcystic-LR of 1.0 ppb (g/L) in drinking water. The U.S. Environmental Protection Agency (EPA) has also established guidelines for Microcyst ins in drinking water:
-For children below school age, 0.3 g/L (ppb)
-For all other age groups, 1.6 g/L (ppb)

2. Introduction

Attogene’s Microcystic Lateral Flow Kit can be used to detect Microcyst ins in liquid samples.

Format: 10 test cassettes, Run Time: 15 Minutes

The most frequently reported cyanotoxins are the hepatotoxic Microcystins (MCs). MCs are peptides with a molecular weight ranging from 900 to 1,100 Da. They consist of seven amino acids of which the two terminal amino acids of the linear peptide are condensed to form a cyclic compound.

A tiered notification system which takes different actions based on different numeric thresholds for Microcytin-LR concentrations in drinking waters has been developed. This is guidance that allows states to take various advisory actions to the public. Our test is a rapid screening tool that enables more comprehensive sampling and aid with rapid decisions concerning actions to take to protect public health.

3. Kit Contents

Attogene AU2024-02 Microcystin Lateral Flow Kit User Manual - Kit
Contents

4. Storage and Stability

The kit should be stored at 2 – 30 until ready to use.
The test must remain in the sealed pouch until use.

5. Required Materials Not Supplied

Timer – For timing use
Centrifuge – For preparation of clear specimens
200ul Pipettor and pipette tips to transfer samples and controls

6. Precautions

The Microcystic Lateral Flow Kit provide only preliminary qualitative test results. Use another, more quantitative, analytical method such as ELISA or instrumental analysis to obtain a confirmed quantitative analytical result. Attogene microcystin ELISA kit can be used for this purpose (catalog number EL2024).
Prior to use, ensure that the product has not expired by verifying that the date of use is prior to the expiration date on the label.
The test cassettes are individually packaged in a foil pouch with a desiccant.
Avoid cross-contamination of samples by using a new tube and disposable pipette tip for each
sample.
Use only Microcystic Lateral Flow Kit reagents from one kit lot, as they have been adjusted in combination.
It is good laboratory practice to use positive and negative controls to ensure proper test performance. Samples which do not contain microcystins (negative controls) as well as samples containing known quantities of microcystins (positive controls) should be analysed with each lot of test strips to provide a reference for line intensity to be expected.

7. Water Collection and Storage

Collect samples into a Sample Bottle and store refrigerated for up to 5 days. If samples must be held for greater than 5 days, samples should be stored frozen.
Allow the test cassettes, running buffer, and samples to reach room temperature before testing.

8. Procedure

Open two sealed cassette. It is recommended to perform steps A and B for each sample evaluation.

A. Sample:
NOTE: Each sample dilution buffer tube contains 20ul of a buffering solution necessary for proper running of the strip. Perform a quick centrifugation of the sample dilution buffer tube to ensure the buffer solution is at the bottom of the tube and not stuck on the sides or top.
o Using the transfer pipet, add 200ul of sample into to one of the sample dilution buffer tubes.
o Mix the sample with the sample buffering solution by pipetting up and down 3 times. o Using a pipet add the buffered sample directly into the sample port of the cassette.

B. Negative Control:
o Pipet 200ul of the solution in the Negative Control tube directly into the sample port of the cassette.

C. Immediately following steps, A and B
o Set a timer for 15 minutes.
o Read results using your eye or with a lateral flow reader. For additional details see section 9 on interpretation of results.

9. Interpretation of Results

This test is a competitive lateral flow assay containing a control line dependent on the amount of microcystins is in the sample. What to expect at the test line: The higher the concentration of microcystin is in the sample the lower the intensity of the test line compared to the negative control. What to expect at the control line: The intensity of the control generally will increase as the test line decreases.

For samples prepared as described above, screening concentrations are determined by comparison of the intensity of the test line on parallel test strips preferably using a lateral flow reader. Test strips with a test line which is darker than or of equal intensity to the test line of the control indicates a result which is the sample is negative or below the limit of detection of the test. Test strips with a test line which is lighter than the control strip indicates a result which is 0.1 ppb. Test strips with no test line visible (only the control line is visible) indicates a result which is 5 ppb. Results should be determined within 15 minutes after completion of the strip test procedure. Determination made using strips which have dried for more or less than the required time may be inaccurate, as line intensities may vary with drying time.

Attogene AU2024-02 Microcystin Lateral Flow Kit User Manual - Microcystin
concentration

The appearance of test strips may also be compared to the illustration on the left to determine approximate sample concentration ranges however, for higher level of confidence a lateral flow reader should be utilized. Please note that the illustration is intended for the demonstration of test line to control line intensity only in running buffer spiked with the indicated amount of microcystin LR. Results should not be determined by comparing the intensity of test lines from test strips to the test line intensity of the illustration, as the overall intensity of test strips may vary slightly with different lots of reagents. To obtain semi-quantitative results in the range of 0.1 5 ppb, solutions of known Microcystin concentration (control solutions) must be tested concurrently with samples. Sample test line intensities can then be compared with control solution test line intensities, yielding approximate sample concentrations. Do not use strips run previously to determine semi- quantitative sample concentrations, as test line intensities may vary once strips are completely dry.

10. Additional Analysis

If necessary, positive samples can be confirmed by ELISA, HPLC, or other conventional methods. A lateral flow reader may also be employed to generate numerical readings from the visual result.

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Attogene is a biotechnology company located in Austin, Texas. Our focus is to enhance health and wellness by offering and developing customer focused Life Science Products domestically and internationally. Our mission is to: