[Questions]{.underline}

[Testing for Aflatoxin]{.underline}

**A**. **[General Questions]{.underline}**

1\. Retest or appeal inspection testing performed under the AMA must be
based on the official file sample.

\(a\) True (b) False

2\. What is the minimum size required for the testing of official
aflatoxin on a barge or sublot?

\(a\) 5 pounds (c) 15 pounds

\(b\) 10 pounds (d) None of the above

3\. Aflatoxin is a naturally occurring mycotoxin produced by two types
of mold known as Aspergillus flavus and Aspergillus parasiticus.

\(a\) True (b) False

4\. Qualitative testing for aflatoxin involves:

\(a\) Running ground corn through a 1/20 sieve.

\(b\) Measuring the level of aflatoxin against a set threshold such as
20 ppb.

\(c\) Measuring the level of aflatoxin in ppb.

(d) All of the above.

5\. Quantitative testing provides aflatoxin results in actual ppb.

\(a\) True (b) False

6\. What is the minimum sample size required for the testing of official
samples of aflatoxin on trucks?

\(a\) 2 pounds (c) 5 pounds

\(b\) 1 pound (d) None of the above

7\. What is the minimum size required for a representative file sample?

\(a\) 250 grams (c) 100 grams

\(b\) 500 grams (d) 1,000 grams

8\. Review inspection testing performed under the USGSA must be based on
the official file sample.

\(a\) True (b) False

9\. What is the minimum sample size required for the official testing of
aflatoxin on railcars?

\(a\) 5 pounds (c) 2 pounds

\(b\) 3 pounds (d) None of the above

10\. When requesting aflatoxin testing service on export shiplots of
grain, the applicant must specify on the load order or application for
inspection:

\(a\) The maximum acceptable aflatoxin limit if different than 20 ppb.

\(b\) The testing methodology desired (qualitative or quantitative).

\(c\) If the grade and aflatoxin are reported on one certificate or
separate certificates.

\(d\) All of the above

11\. Quantitative testing provides aflatoxin results in actual ppb.

\(a\) True (b) False

12\. Grains other than corn may be tested for aflatoxin.

\(a\) True (b) False

13\. What are the two types of aflatoxin testing services provided by
official personnel?

(a) Qualitative and HPLC

(b) Black Light and Quantitative

(c) Veratox and Aflatest

\(d\) Qualitative and Quantitative

14\. What is the recommended or optimum sample size required for testing
of official samples?

\(a\) 5 pounds (c) 15 pounds

\(b\) 10 pounds (d) None of the above

15\. Board appeal inspections for aflatoxin are limited to the analysis
of a file sample.

\(a\) True (b) False

16\. All appeal samples will be based on the official file sample using:

\(a\) Qualitative methodology.

\(b\) Quantitative methodology.

\(c\) Applicant choice.

\(d\) High Performance Liquid Chromatography (HPLC) only

**B. [Vicam Aflatest Specific Questions]{.underline}**

1\. Each fluorometer is equipped with a set of standard vials packaged
three vials per case which must be:

\(a\) Refrigerated when not in use.

\(b\) Stored in a cabinet or drawer in the original case when not in
use.

\(c\) Placed under a fume hood before, during, and after use.

\(d\) None of the above.

2\. During calibration, before placing standard vials in the cuvette
chamber, you should:

\(a\) Wipe each of the vials with a clean cloth or paper wipe.

\(b\) Wash each vial with a bleach solution.

\(c\) Set the calibration value at 20.0 ppb.

\(d\) None of the above.

3\. The extraction method used for Aflatest (testing for corn) includes
adding 100 mL of 80/20 methanol/water mixture to 50 grams of ground
sample and 5 grams of salt, then blending the mixture for exactly 1
minute.

\(a\) True (b) False

4.  When checking the calibration (for testing corn) of the Series IV
    fluorometer with the yellow vial, the value displayed after it has
    been fully inserted into the sample well should be 78 ±4 ppb.

> \(a\) True (b) False

5\. Once prepared the dilute developer solution can be used for no more
than 12 hrs.

\(a\) True (b) False

6\. Mix 5 ml. of developer solution with \_\_\_\_\_\_\_\_\_\_ mL of
distilled water to prepare the dilute developer solution.

\(a\) 45

(b) 5

(c) 100

(d) 9

7\. How often is the calibration checked using the yellow vial?

\(a\) Every 4 hours (c) Every 3 hours

\(b\) Every 2 hours (d) Every hour

8\. When performing solution testing, a combination of 1 mL dilute
developer and 1 mL methanol should read between -3.0 and
\_\_\_\_\_\_\_\_\_.

\(a\) +2.0 (c) +3.0

\(b\) +1.0 (d) +4.0

9\. Aflatest is a qualitative test method that provides test results in
actual ppb.

\(a\) True (b) False

10\. Aflatest may be used as a qualitative method to report results as
being equal to or exceeding a threshold.

\(a\) True (b) False

11\. The dilute developer solution and HPLC grade methanol must be
checked for \_\_\_\_\_\_\_\_\_\_\_\_\_\_\_ after calibrating the
fluorometer.

(a) Nothing

(b) All reagents should be giving a positive reaction

(c) Background fluorescence

(d) Reddish brown color

**C. [Neogen AgriScreen Specific Questions]{.underline}**

1\. The Agri‑Screen test is an enzyme-linked immunoassay test that
provides easy‑to‑read visual color results.

\(a\) True (b) False

2\. Each cellophane bag containing components of the kit is marked with
a label indicating the lot number and expiration date.

\(a\) True (b) False

3\. The green‑capped tubes (TMB) in the substrate package are sensitive
to the light. If the contents have a \_\_\_\_\_\_\_\_\_ do not use the
tube, discard and select another tube.

(a) Blue color

(b) Red color

(c) Clear color

(d) Yellow color

4\. If the sample results are a darker (bluer) or the same as the
control well the results should be considered to be negative or ≤ 20
ppb.

\(a\) True (b) False

5\. Sample extraction procedures require that:

\(a\) 50 grams of ground sample be mixed with 100 ml of 55%
methanol/water

solution and blended on high speed for one minute.

> \(b\) 50 grams of ground sample be mixed with 250 ml of 80%
> methanol/water
>
> solution and blended on high speed for three minutes.
>
> \(c\) 50 grams of ground sample be placed in a whirlpak with 250 ml of
> 70%
>
> methanol/water solution and mixed by shaking by hand for 3 minutes.
>
> \(d\) None of the above.

6\. How long will the substrate solution last if kept in the dark?

\(a\) One day (c) Three days

> \(b\) One week (d) Five days

7\. There are two well strips used in the test. The strip with the red
marked line is called the mixing well strip.

\(a\) True (b) False

8\. A preset amount of liquid is the amount drawn into the pipette tip
when the syringe plunger is depressed and then released slowly.

\(a\) True (b) False

9\. To wash out the well once the initial reaction is completed,

\(a\) Shake out the contents of the wells.

> \(b\) Fill each well with distilled water using a wash bottle.

\(c\) Remove all water droplets by turning wells upside down and

> vigorously tap wells on a paper towel.
>
> \(d\) All of the above

10\. Agri‑Screen kits have to be refrigerated.

\(a\) True (b) False

11\. The green‑capped tubes (TMB) in the substrate package are sensitive
to the light. If the contents have a blue color, do not use the tube,
discard and select another tube.

\(a\) True (b) False

12. There are two well strips used in the test. The plain well strip is
    called the anti-body coated well strip.

\(a\) True (b) False

**D. [Neogen Veratox AST Specific Questions]{.underline}**

1\. The "overfill method" of pipetting is used for:

(a) Mixing the contents of the mixing wells.

(b) Placing 100 uL of substrate into each well.

(c) Adding 100 uL of red stop solution into each well.

(d) All of the above.

2\. The kit is packaged in a sealed "foil bag" with a label indicating
the lot number and production date.

\(a\) True (b) False

3\. Mix the ground sample and extraction solution by:

(a) Shaking by hand for exactly 2 minutes.

(b) Blending on high speed for exactly 2 minutes.

(c) Shaking by hand or blending for exactly 2 minutes.

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(d) Shaking by hand or blending for exactly 3 minutes.

4\. How many mixing wells are used for each sample?

(a) One for the sample portion.

(b) Two -- one control and one sample portion.

(c) Three -- one control and two sample portions.

(d) Four -- two controls and two sample portions.

5\. The Veratox AST measures total aflatoxins (B-1, B-2, G-1, G-2) in
corn, soybeans,

> wheat, sorghum and certain other commodities using an enzyme-linked
> immunosorbent assay (ELISA) technique.
>
> \(a\) True (b) False

6\. Sample extraction procedures require that:

\(a\) 50 grams of ground sample be mixed with 100 ml of 55%
methanol/water

> solution and blended on high speed for one minute.
>
> \(b\) 50 grams of ground sample be mixed with 250 ml of 80%
> methanol/water
>
> solution and blended on high speed for three minutes.
>
> \(c\) 50 grams of ground sample be mixed with 250 ml of 70%
> methanol/water solution and blended on high speed for exactly 2
> minutes.
>
> \(d\) None of the above.

7\. To wash out the antibody wells once the initial reaction is
completed,

\(a\) Shake out the contents of the wells.

> \(b\) [.]{.underline} With a wash bottle containing
> deionized/distilled water, fill each antibody well and dump the
> contents into a waste receptacle. Repeat this step 5 times
>
> \(c\) With a wash bottle containing deionized/distilled water, fill
> each antibody well and dump the contents into a waste receptacle.
> Repeat this step 15 times.
>
> \(d\) Turn microwell holder, with wells in it, upside down on a paper
> towel and tap gently until the solution is removed from the wells.

8\. When not in use, test kits must be stored:

(a) In a freezer.

(b) At room temperature.

> \(c\) Refrigerated at temperatures between 36°- 46° F.

\(d\) Either (b) or (c).

9\. Mixing in the antibody-coated wells is accomplished by:

(a) Vigorously sliding the microwell holder back and forth on a
    horizontal surface for 5 seconds.

(b) Vigorously sliding the microwell holder back and forth on a
    horizontal surface for 15 seconds.

(c) Gently sliding the microwell holder back and forth on a horizontal
    surface for 15 seconds.

(d) Gently sliding the microwell holder back and forth on a horizontal
    surface for 5 seconds.

10\. How many sample portions are tested for each sample?

(a) One (c) Three

> \(b\) Two (d) Four

11\. The Micro-well reader is designed to do several testing functions
with each function having specific set-up parameters. If the display set
up code of the reader is different than the FGIS specified code, the
operator must:

(a) Disregard the error and proceed to run samples.

(b) Contact the Neogen Corporation representative for instruction.

(c) Use a different test kit.

> \(d\) Contact the field office for instruction.

**E. [Romer Fluoroquant FQSpecific Questions]{.underline}**

1\. The Romer Fluoroquant aflatoxin test method allows quantitative
measurement of total aflatoxins (B1, B2, G1, and G2) in parts per
billion for corn, corn meal, rice, popcorn, sorghum, and wheat.

> \(a\) True (b) False

2\. To prepare the working developer reagent add of deionized or
distilled water to the bottle supplied with the repipettor, and then add
the contents of one ampule of developer concentrate.

\(a\) 50 mL

(b) 100 mL

(c) 250 mL

(d) 10 mL

3\. In preparation of the developer solution, how many times must the
developer concentrate ampule be rinsed with the working developer
solution?

\(a\) One time (c) Five times

\(b\) Three times (d) Two times

4\. The developer working reagent must be made fresh every \_\_ hours.

(a) 24

(b) 16

\(c\) 8

(c) 2

5\. To insure a complete extract solution purification, the solution
must be pushed through the column slowly until air comes out the bottom.
What is the time frame recommended for completing this step?

\(a\) At least 20 - 30 seconds

\(b\) No more than 20 - 30 seconds

\(c\) At least 30 - 40 seconds

\(d\) No more than 30 - 40 seconds

6\. In the purification process for corn, corn meal, rice, popcorn,
sorghum, and wheat, how much sample extract is placed into the top of
the column?

\(a\) 0.5 mL (c) 1.0 mL

\(b\) 1.0 uL (d) 2.0 mL

7\. In the purification process for corn, corn meal, rice, popcorn,
sorghum, and wheat, sample extract and diluent are added to the column,
what is the quantity of diluent added to the column?

\(a\) 1.0 mL (c) 0.5 mL

\(b\) 1.0 uL (d) 0.5 uL

8\. In the purification process, after the sample extract and diluent
are added to the column, the column is capped and mixed by hand shaking.
How long should the column be shaken?

\(a\) 10 seconds (c) 15 seconds

\(b\) 1 minute (d) 5 seconds

9\. In the derivatization process, what is the quantity of developer
working reagent added to each purified sample?

\(a\) 0.5 mL (c) 1.5 mL

\(b\) [1.0 mL]{.underline} (d) 1.0 uL

10\. What is the maximum number of samples that may be derivatized at
one time?

\(a\) One (c) Up to five

\(b\) Two (d) As many as you can possibly do

11\. UniSep 2001 columns and calibrators must be stored at room
temperature.

\(a\) True (b) False

12. To test the control standard use the wheat calibration factors.

> \(a\) True (b) False

13\. Describe the storage conditions for the positive control standard.

\(a\) Room temperature in a sealed container.

\(b\) A cool, dry, well ventilated area away from heat sources.

> \(c\) A cool, dry, well ventilated area away from direct sunlight,
> combustible materials, and incompatible substances.
>
> \(d\) Refrigerated at 23° to 32° F

14\. Sample extraction procedures for the Fluoroquant test for corn
require that:

\(a\) 50 grams of ground sample is mixed with 100 mL of 80%
methanol/water

> solution and blended on high speed for one minute.
>
> \(b\) 50 grams of ground sample is mixed with 250 mL of 80%
> methanol/water

solution and blended on high speed for three minutes.

> \(c\) 50 grams of ground sample is placed in a whirlpak with 250 mL of
> 70%

methanol/water solution and mixed by shaking by hand for 3 minutes.

> \(d\) None of the above.

15\. Sample extraction procedures for the Fluoroquant test on soybeans
and corn/soy blend require that:

\(a\) 50 grams of ground sample is mixed with 100 mL of 80%
methanol/water

solution and blended on high speed for one minute.

> \(b\) 50 grams of ground sample is mixed with 250 mL of 80%
> methanol/water

solution and blended on high speed for three minutes.

> \(c\) 50 grams of ground sample is placed in a whirlpak with 100 mL of
> 90%
>
> acetonitrile/water solution and blended on high speed for one minute.

(d) None of the above.

**F. [SDI Myco ]{.underline}** **[Specific Questions]{.underline}**

1\. Sample extraction procedures require that:

\(a\) 50 grams of ground sample be mixed with 100 ml of 55%
methanol/water

> solution and blended on high speed for one minute.
>
> \(b\) 50 grams of ground sample be mixed with 250 ml of 80%
> methanol/water
>
> solution and blended on high speed for three minutes.
>
> \(c\) 50 grams of ground sample be mixed with 250 ml of 70%
> methanol/water solution and blended on high speed for exactly 2
> minutes.
>
> \(d\) None of the above.

2\. What is the maximum number of test samples that can be run at one
time?

\(a\) two (c) six

\(b\) twelve (d) nineteen

3\. How many sample portions are tested for each sample?

> \(a\) One (c) Three
>
> \(b\) Two (d) Four

4\. When not in use, test kits must be stored:

(a) In a freezer.

(b) At room temperature.

\(c\) At temperatures between 36° - 46° F.

(c) Either (b) or (c).

5\. The Myco  test is a competitive enzyme-linked immunosorbent assay
that provides quantitative measurement for the presence of aflatoxin in
corn, corn meal, corn soy blend sorghum and popcorn.

\(a\) True (b) False

6\. The extraction solvent used in the Myco  test method is a
methanol/water (distilled or deionized) mixture consisting of 70 percent
methanol (ACS grade or better) and

> 30 percent distilled or deionized water.

\(a\) True (b) False

7\. After the reaction mixture incubates for 5 minutes, dump the
contents of the wells into a waste container and wash each well by
vigorously overfilling. Repeat the vigorous wash for a total of
\_\_\_\_\_ washes.

\(a\) two (c) three

\(b\) four (d) five

8\. What instrument is used to read the optical density of test wells?

\(a\) Fluorometer (c) Stat-Fax microwell reader

\(b\) EL-301 microwell reader (d) Hyperion MicroReader 3 well reader

9.  To prepare the Wash Solution transfer the contents of the Wash
    Concentrate into a

500-ml plastic bottle and add \_\_\_\_\_\_ ml of distilled or deionized
water.

\(a\) 200 (c) 4750

\(b\) 475 (d) 1000

10\. After the substrate solution is added to the test well(s), incubate
for:

\(a\) exactly five minutes (c) five minutes

\(b\) exactly ten minutes (d) none of the above

11\. Standard certification procedures require test results exceeding 80
ppb to be reported as \> 80 unless a supplemental analysis is performed.

\(a\) True (b) False

**G.** **[R -Biopharm Ridascreen FAST Aflatoxin SC- Specific
Questions]{.underline}**

1\. The extraction solvent used for the Ridascreen test is made up by
using the ratio of 70 parts of methanol to 30 parts of water.

\(a\) distilled (c) distilled or deionized

\(b\) deionized (d) tap

2\. What is the size of ground portion that is used for testing?

\(a\) 50 grams (c) 100 grams

\(b\) 250 grams (d) one pound

3\. In the extraction process, 250 ml of the extraction solvent is added
to the ground portion and blended for minutes.

\(a\) one (c) three

\(b\) two (d) five

4\. Following the extraction process, filter ml of the extract through a
filtering syringe.

\(a\) 1 (c) 2.0

\(b\) 1.5 (d) 2.5

5\. To analyze a single sample, use one red-marked mixing well for the
test sample and

wells for controls.

\(a\) one (c) two

\(b\) three (d) five

6\. What is the maximum number of samples that can be tested at one
time, per set of control standards?

\(a\) 19 (c) 21

\(b\) 7 (d) 23

7\. After adding the sample solution, enzyme conjugate, and
anti-aflatoxin antibody to the wells, mix the wells by gently sliding
the plate back and forth on a flat surface. After mixing, then incubate
for minutes.

\(a\) ten (± 0.5 minutes) (c) five (± 0.5 minutes)

\(b\) ten (± 1.0 minutes) (d) fifteen (± 0.5 minutes)

8\. Which microwell reader(s) is/are used for reading test results?

\(a\) Biotek EL-301 (c) Stat-Fax Model 303 PLUS

\(b\) Hyperion Model 4027-002 (d) Both (a) and (c)

9\. Test results should be read within minutes of completion of the
test.

\(a\) five (c) ten

\(b\) twenty (d) thirty

10\. Standard certification procedures require test results exceeding 50
ppb to be reported as \> 50 unless a supplemental analysis is performed.

\(a\) True (b) False

11\. Test kits should be refrigerated at temperatures between 35°F and
46°F.

> \(a\) True (b) False

12\. A value of less than 0.6 absorbance units for the zero standard may
indicate deterioration of reagents.

\(a\) True (b) False

**H.** **[Envirologix QuickTox- Specific Questions]{.underline}**

1\. The QuickTox test kit uses lateral flow test strip technology that
provides quantitative results.

\(a\) True (b) False

2\. An extraction solvent used for the QuickTox test is made up by using
the ratio of 70 parts of methanol to 30 parts of water.

\(a\) distilled (c) distilled or deionized

\(b\) deionized (d) tap

3\. What is the size of ground portion that is used for testing?

\(a\) 50 grams (c) 100 grams

\(b\) 250 grams (d) one pound

4\. In the extraction process, 100 ml of the extraction solvent is added
to the ground portion and shaken (by hand) for minutes.

\(a\) one (c) three

\(b\) two (d) five

5\. If a mechanical shaker is used to mix the extraction solvent and
ground sample portion shaking time may be reduced to:

\(a\) two minutes (c) 1.5 minutes

\(b\) one minute (d) 30 seconds

6\. After shaking the sample, the extraction portion will separate into
2 layers. The top (yellowish) layer containing the aflatoxin residues
will be used for the testing.

\(a\) True (b) False

7\. To prepare the reaction vial, place 150 µl distilled/deionized water
into the vial.

\(a\) True (b) False

8\. Test strips are placed into the reaction vial containing the diluted
sample extract. The arrow tape on the end of the strip should point into
the reaction vial.

\(a\) True (b) False

9\. Allow the test strip to develop for minutes before making final
assay interpretations.

\(a\) two minutes (b) two to three minutes

\(b\) five minutes (d) ten minutes

10\. Development of a Control Line within 5 minutes indicates that the
strip functioned properly.

\(a\) [True]{.underline} (b) False

11\. A sample containing aflatoxin residues more than Level of Detection
will develop 2 distinct lines, the Control Line and the Test Line, in
the test area.

\(a\) True (b) False

12\. Test kits should be refrigerated at temperatures between 36°F and
48°F.

> \(a\) True (b) False

**I.** **[R-Biopharm AFLACARD T20 -- Specific Questions]{.underline}**

1.  The AFLACARD T20 test kit uses an enzyme immunoassay procedure that
    provides qualitative results.

> \(a\) True (b) False

2.  Kits should be brought to room temperature (68 º - 82 ºF) prior to
    use. This will take approximately 30 minutes.

> \(a\) True (b) False

3.  The extraction solvent used for the AFLACARD T20 test is made up by
    using the ratio of 80 parts of methanol to 20 parts of
    \_\_\_\_\_\_\_\_\_ water.

> \(a\) distilled (c) distilled or deionized
>
> \(b\) deionized (d) tap

4.  What is the size of ground portion that is used for extraction?

> \(a\) 50 grams (c) 100 grams
>
> \(b\) 250 grams (d) one pound

5.  In the extraction process, 100 ml of the extraction solvent is added
    to the ground portion and blended on high speed for \_\_\_
    minute(s).

> \(a\) one (c) three
>
> \(b\) two (d) five

6.  After extraction, filter through a Whatman No. 4 filter paper and
    collect a minimum of 10 ml of the extract into a sample jar labeled
    with the sample identification.

> \(a\) True (b) False

7.  The filtrate in the sample jar must be diluted. Remove the lid from
    the sample diluent tube and add \_\_\_\_\_\_\_ of filtrate.

> \(a\) 1 ml (c) 200 µl
>
> \(b\) 250 µl (d) 3 drops

8.  When applying diluted filtrate (or any other liquid) to the test
    port, always ensure that the liquid had passed completely through
    the membrane before proceeding to the next step.

> \(a\) True (b) False

9.  The control spot must not develop any detectable color in order to
    have a valid test result.

> \(a\) True (b) False

10. The sample should be considered to be negative when the sample and
    control spot both have clearly visible color development.

> \(a\) True (b) False

11. For both negative and positive results, dispose of any remaining
    liquid filtrate in the chemical waste container.

> \(a\) True (b) False

12. For long term storage, test kits may be frozen.

> \(a\) True (b) False

**J [Neogen REVEAL FOR AFLATOXIN -- Specific Questions]{.underline}**

1.  The REVEAL FOR AFLATOXIN test kit uses a lateral flow strip
    procedure that provides qualitative results.

> \(a\) True (b) False

2.  Kits should be at room temperature (64 º - 86 ºF) prior to use.

> \(a\) True (b) False

3.  The extraction solvent used for the REVEAL FOR AFLATOXIN test is
    made up by using the ratio of 70 parts of methanol to 30 parts of
    \_\_\_\_\_\_\_\_\_ water.

> \(a\) distilled (c) distilled or deionized
>
> \(b\) deionized (d) tap

4.  What is the size of ground portion that is used for extraction?

> \(a\) 50 grams (c) 100 grams
>
> \(b\) 250 grams (d) one pound

5.  In the extraction process, 100 ml of the extraction solvent is added
    to the ground portion and blended on high speed for \_\_\_
    minute(s).

> \(a\) one (c) three
>
> \(b\) two (d) five

6.  After extraction, allow the sample to settle, filter, and collect
    the extract in a sample jar labeled with the sample identification.

> \(a\) True (b) False

7.  Pipette 200 µl of sample diluent to a sample cup. Using a new tip,
    pipette \_\_\_\_\_\_\_ of the filtered extract into the cup. Mix the
    solution by pipetting up and down 3 times.

> \(a\) 1 ml (c) 200 µl
>
> \(b\) 250 µl (d) 3 drops

8.  Allow the strip to develop in the sample cup for 5 minutes

> \(a\) True (b) False

9.  A strip developing a detectable line in the Control Zone indicates
    that the strip has functioned properly.

> \(a\) True (b) False

10. The sample should be considered to be negative when the test zone
    and control zone both have clearly visible lines present.

> \(a\) True (b) False

11. For both negative and positive results, dispose of any remaining
    liquid filtrate in the chemical waste container.

> \(a\) True (b) False

12. For long term storage, test kits may be frozen.

> \(a\) True (b) False

K.  **[Romer AGRASTRIP TEST KIT -- Specific Questions]{.underline}**

    1.  The AGRASTRIP test kit uses a lateral flow strip procedure that
        provides qualitative results.

> \(a\) True (b) False

2.  Store kit components at 35 º - 86 ºF when not in use to assure full
    shelf life.

> \(a\) True (b) False

3.  The extraction solvent used for the AGRASTRIP test is made up by
    using the ratio of 70 parts of methanol to 30 parts of
    \_\_\_\_\_\_\_\_\_ water.

> \(a\) distilled (c) distilled or deionized
>
> \(b\) deionized (d) tap

4.  What is the size of ground portion that is used for extraction?

> \(a\) 50 grams (c) 100 grams
>
> \(b\) 250 grams (d) one pound

5.  In the extraction process, 100 ml of the extraction solvent is added
    to the ground portion and blended on high speed for \_\_\_
    minute(s).

> \(a\) one (c) three
>
> \(b\) two (d) five

6.  After blending, filter the extract through filter paper into a
    sample jar labeled with the sample identification.

> \(a\) True (b) False

7.  Pipette 50 µl of assay diluent to a sample microwell. Dissolve the
    coating by pipetting up and down 5 times. Using a new tip, pipette
    \_\_\_\_\_\_\_ of the sample extract into the microwell. Mix the
    solution by pipetting up and down 3 times.

> \(a\) 1 ml (c) 200 µ[l]{.underline}
>
> \(b\) 50 µl (d) 3 drops

8.  Allow the strip to develop in the sample microwell for 5 minutes

> \(a\) True (b) False

9.  A strip developing a detectable control line indicates that the
    strip has functioned properly.

> \(a\) True (b) False

10. The sample should be considered to be negative when a test line and
    control line are both clearly visible.

> \(a\) True (b) False

11. For both negative and positive results, dispose of any remaining
    liquid filtrate in the chemical waste container.

> \(a\) True (b) False

12. For long term storage, test kits may be frozen.

> \(a\) True (b) False

**L. [Neogen Veratox Specific Questions]{.underline}**

> 1\. The extraction solvent used for the Veratox test is made up by
> using the ratio of 70 parts of methanol to 30 parts of
> \_\_\_\_\_\_\_\_\_ water.

\(a\) distilled (c) distilled or deionized

> \(b\) deionized (d) tap
>
> 2\. What is the size of ground portion that is used for extraction?
>
> \(a\) 50 grams (c) 100 grams
>
> \(b\) 250 grams (d) one pound
>
> 3\. Mix the ground sample and extraction solution by:

(a) Shaking by hand for exactly 2 minutes.

(b) Blending on high speed for exactly 2 minutes.

(c) Shaking by hand or blending for exactly 1 minute.

(d) Shaking by hand or blending for exactly 3 minutes.

> 4\. The antibody wells are packaged in a sealed "foil pack" with
> desiccant.
>
> \(a\) True (b) False
>
> 5\. How many mixing wells are used for a single sample?

(a) Two -- one control and one sample portion.

(b) Three -- one control and two sample portions.

(c) Four -- two controls and two sample portions.

(d) Five -- Four controls and one sample portion.

> 6\. What is the maximum number of test samples that can be run at one
> time?

\(a\) two (c) six

> \(b\) twelve [(]{.underline}d) twenty
>
> 7\. Mixing in the antibody-coated wells is accomplished by:

(a) Vigorously sliding the microwell holder back and forth on a
    horizontal surface for 5 seconds.

(b) Vigorously sliding the microwell holder back and forth on a
    horizontal surface for 15 seconds.

(c) Gently sliding the microwell holder back and forth on a horizontal
    surface for 10 to 20 seconds.

(d) Gently sliding the microwell holder back and forth on a horizontal
    surface for 5 seconds.

> 8\. To wash out the antibody wells once the initial reaction is
> completed,
>
> \(a\) Shake out the contents of the wells.
>
> \(b\) With a wash bottle containing deionized/distilled water, fill
> each antibody well and dump the contents into a waste receptacle.
> Repeat this step 5 times.
>
> \(c\) With a wash bottle containing deionized/distilled water, fill
> each antibody well and dump the contents into a waste receptacle.
> Repeat this step 15 times.
>
> \(d\) Turn microwell holder, with wells in it, upside down on a paper
> towel and tap gently until the solution is removed from the wells.
>
> 9\. After the addition of substrate, mix and allow the microwells to
> incubate for 5 minutes.
>
> \(a\) True (b) False
>
> 10\. Results should be read within 20 minutes of completion of the
> test.
>
> \(a\) True (b) False
>
> 11\. When not in use, test kits must be stored:

(a) in a freezer.

(b) At room temperature.

> \(c\) Refrigerated at temperatures between 36°- 46° F.
>
> \(d\) Either (b) or (c).
>
> 12\. For both negative and positive results, decontaminate lab ware
> with a bleach solution.
>
> \(a\) True (b) False

**M. [Charm Science ROSA® P/N Specific Questions]{.underline}**

1.  The ROSA® P/N test kit uses a lateral flow strip procedure that
    provides qualitative results.

> \(a\) True (b) False

2.  Test strips should be at room temperature (64 º - 86 ºF) prior to
    use.

> \(a\) True (b) False

3.  The extraction solvent used for the ROSA® P/N test is made up by
    using the ratio of 70 parts of methanol to 30 parts of
    \_\_\_\_\_\_\_\_\_ water.

> \(a\) distilled (c) distilled or deionized
>
> \(b\) deionized (d) tap

4.  What is the size of ground portion that is used for extraction?

> \(a\) 50 grams (c) 100 grams
>
> \(b\) 250 grams (d) one pound

5.  In the extraction process, 100 ml of the extraction solvent is added
    to the ground portion and shaken 30 seconds and allowed to settle
    for \_\_\_\_\_ minute(s).

> \(a\) one (c) three
>
> \(b\) two (d) five

6.  Store the Positive and Negative controls at 32 - 45 º F for up to
    one week.

> \(a\) True (b) False

7.  Pipette \_\_\_\_\_ µl of clarified extract to a labeled
    micro-centrifuge tube containing a 1.0 ml. AFQ Dilution Buffer
    and mix. This is the diluted extract.

> \(a\) 1 ml (c) 100 µl
>
> \(b\) 250 µl (d) 3 drops

8.  Allow the strip to incubate in the Quad incubator for 5 minutes

> \(a\) True (b) False

9.  A test is invalid if a Control Line is missing, smeared or uneven,
    or, if the Test Line is uneven.

> \(a\) True (b) False

10. A sample containing aflatoxin residues in excess of 20 ppb will
    develop a Test Line that is lighter in intensity than the Control
    Line.

> \(a\) True (b) False

11. The LF-Rosa Reader-M can be used to interpret the strip and display
    the results.

> \(a\) True (b) False

12. Certify results as being equal to or less than 20 ppb or exceeding
    20 ppb, as applicable.

> \(a\) True (b) False

**N. [Charm Science ROSA® Aflatoxin (Quantitative) Specific
Questions]{.underline}**

1.  The ROSA® Quantitative test kit uses a lateral flow strip procedure
    that provides qualitative results.

> \(a\) True (b) False

2.  Test strips should be at room temperature (64 º - 86 ºF) prior to
    use.

> \(a\) True (b) False

3.  The extraction solvent used for the ROSA® P/N test is made up by
    using the ratio of 70 parts of methanol to 30 parts of
    \_\_\_\_\_\_\_\_\_ water.

> \(a\) distilled (c) distilled or deionized
>
> \(b\) deionized (d) tap

4.  What is the size of ground portion that is used for extraction?

> \(a\) 50 grams (c) 100 grams
>
> \(b\) 250 grams (d) one pound

5.  In the extraction process for soybeans, \_\_\_\_\_\_\_ ml of the
    extraction solvent is added to the ground portion and shaken 30
    seconds and allowed to settle for one minute(s).

> \(a\) 150 ml. (c) 50 ml.
>
> \(b\) 100 ml. (d) 200 ml.

6.  Store the Positive and Negative controls at 32 - 45 º F for up to
    one week.

> \(a\) True (b) False

7.  Pipette \_\_\_\_\_ µl of clarified extract to a labeled
    micro-centrifuge tube containing a 1.0 ml. AFQ Dilution Buffer
    and mix. This is the diluted extract.

> \(a\) 1 ml (c) 100 µl
>
> \(b\) 250 µl (d) 3 drops

8.  Allow the strip to incubate in the Quad incubator for 10 minutes

> \(a\) True (b) False

9.  Positive control must read between 20 -- 30 ppb.

> \(a\) True (b) False

10. Incubator must be clean, level and at 45 ±1º C.

> \(a\) True (b) False

11. The LF-Rosa Reader-M can be used to interpret the strip and display
    the results.

> \(a\) True (b) False

12. Certify results to the nearest tenth ppb.

> \(a\) True (b) False

**O.** **[Envirologix QuickTox (Released 2006) - Specific
Questions]{.underline}**

1\. The QuickTox test kit uses lateral flow test strip technology that
provides quantitative results.

\(a\) True (b) False

2\. The extraction solvent used for the QuickTox test is made up by
using the ratio of 70 parts of methanol to 30 parts of distilled or
deionized water or 50 parts \_\_\_\_\_\_\_\_\_\_\_\_ to 50 parts of
distilled or deionized water.

\(a\) acetone (c) ethanol

\(b\) methanol (d) tap water

3\. What is the size of ground portion that is used for testing?

\(a\) 50 grams (c) 100 grams

\(b\) 250 grams (d) one pound

4\. In the extraction process, 100 ml of the extraction solvent is added
to the ground portion and shaken (by hand) for minutes.

\(a\) one (c) three

\(b\) two (d) five

5\. If a mechanical shaker is used to mix the extraction solvent and
ground sample portion shaking time may be reduced to:

\(a\) two minutes (c) 1.5 minutes

\(b\) one minute (d) 30 seconds

6\. After shaking the sample, the extraction portion will separate into
2 layers. The top (yellowish) layer containing the aflatoxin residues
will be used for the testing.

\(a\) True (b) False

7\. To prepare the reaction vial, place 150 µl of tap water into the
vial.

\(a\) True (b) False

8\. Test strips are placed into the reaction vial containing the diluted
sample extract. The arrow tape on the end of the strip should point into
the reaction vial.

\(a\) True (b) False

9\. Allow the test strip to develop for minutes before making final
assay interpretations.

\(a\) two minutes (b) two to three minutes

\(b\) five minutes (d) ten minutes

10\. Development of a Control Line within 5 minutes indicates that the
strip functioned properly.

\(a\) True (b) False

11\. A sample containing aflatoxin residues more than 20 ppb will
develop 2 distinct lines, the Control Line and the Test Line, in the
test area.

\(a\) True (b) False

12\. Test kits should be refrigerated at temperatures between 36°F and
48°F.

> \(a\) True (b) False