Fecal Extraction Device

For In Vitro Diagnostic Use

CLIA Complexity: High

REF 504837 Rx Only

∆Intended Use
The Fecal Extraction Device is a single use tube containing extraction buffer intended for sampling and
extracting human stool specimens and subsequent analysis with the QUANTA Flash Calprotectin CIA,
QUANTA Lite Calprotectin ELISA and QUANTA Lite Calprotectin Extended Range ELISA assays.

Description and Explanation of the Test

The Fecal Extraction Device (FED) acquires the amount of stool necessary to perform the QUANTA Flash
Calprotectin CIA, QUANTA Lite Calprotectin ELISA and QUANTA Lite Calprotectin Extended Range
ELISA assays directly from the primary specimen container instead of weighing the sample.
The device consists of a tube, containing 2.8 mL of extraction buffer, and a stick shaped with seven
grooves for collecting the sample. The upper end of the device is made up of two parts which can be
removed with two separate rotations: the white screw cap (connected to the plastic stick with grooves) is
removed by twisting counter-clockwise. The red lower part (for retaining the excess material) is removed
by twisting clockwise. Once having completed the extraction procedure, remove both the white and red
upper parts. The tube containing the extracted sample is directly compatible with the BIO-FLASH
instrument sample rack.

Precautions
1. Handle stool as potentially infectious material.
2. Adaptation of the extraction device for use with automated sample processors and other automated
systems may yield differences in test results from those obtained using the manual procedure. It is
the responsibility for each laboratory to validate that their automated procedure yields results within
acceptable limits.
3. The device is disposable. Observe all federal, state and local environmental regulations when disposing of waste.
4. This product is For In Vitro Diagnostic Use.

Storage Before Use

Store devices at 2-8°C. Devices can be stored at room temperature for up to 72 hours.

Materials provided
- 100 ea. fecal extraction devices containing 2.8 mL of extraction buffer
- 100 ea. white caps

Additional Materials Required But Not Provided

- Centrifuge capable to reach 1000 – 3000 x g
- Laboratory pipette capable of pipetting 56 μL

Specimen Preparation for Analysis

Sampling procedure for feces of normal consistency
1. Pull out the plastic stick of the device by turning the white screw-cap counter-clockwise.
Attention: Do not remove the red part.
2. Dip the stick in the previously collected stool sample.
3. Repeatedly rotate the stick in the stool sample until all grooves are filled with fecal material.
4. Before placing the stick into the device, remove the stool excess by rotating the stick on the internal wall of the stool
container.
5. Fully insert the stick with the fecal material into the tube containing the extraction buffer. While holding the red part of
tube, turn the cap clockwise until completely closed.
Attention: The red part must remain closed.

Sampling procedure for liquid feces or a consistency which does not allow the use of the stick
1. Using the pipette, aspirate 56 μL of liquid stool.
2. Pull out the plastic stick by turning the white screw-cap counter-clockwise.
3. Pipette the liquid stool sample into the tube containing the extraction buffer.
4. Place the stick (which will not contain stool) back into tube containing the extraction buffer. While holding the red part of the
tube, turn the cap clockwise until completely closed.

Storage of the device after stool sampling procedure

If there is a delay in continuing with the laboratory procedure, the device containing the stool sample can be stored at room
temperature for up to 72 hours.

∆Laboratory procedure
1. Bring samples to room temperature.
2. Vortex the device vigorously for 30-60 seconds in order to homogenize the content. Proceed to step 3 if the grooves are
visually free of stool material, otherwise vortex the device again up to a total of four minutes. After a total vortex time of four
minutes proceed to step 3, regardless if residual stool still remains in the grooves.
3. Remove the upper end of the device (turn the red part clockwise; both the red lower part and the white screw cap will come
off together). Close the tube with the extra cap provided.
4. Centrifuge the extraction device for 10 minutes at 1000 - 3000 x g to remove the possible residuals of fecal material. This is
necessary to prevent damage to the aspirating and dispensing systems of the automated processor.
5. The fecal extract is now ready for testing according to the package insert of the QUANTA Flash Calprotectin CIA, QUANTA
Lite Calprotectin ELISA and QUANTA Lite Calprotectin Extended Range ELISA assays.

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Storage of the Fecal Extract
Upon completion of the extraction procedure described above, the fecal extract is stable under the following conditions:
- Up to 72 hours at room temperature
- Up to 14 days at 2-8°C
- Up to 90 days at -20°C. Frozen samples can stand up to 4 freeze/thaw cycles*.
* Thawed samples must be homogenized before its use. Follow steps 2 and 4 of the Laboratory Procedure.

Limitations of the Procedure

1. The performance characteristics have not been established in matrices other than stool.
2. Calprotectin in stool may not be evenly distributed throughout the sample. Without proper homogenization of the sample,
variations can occur.1

Performance Characteristics: Stool Collection

The stool mean volume obtained by using the extraction device is 56 mg. This value has been defined by means of three
methods:
1. Volume calculated for each groove
2. Weighing the plastic stick (including the cap) before and after sampling: Five different stool samples encompassing various
stool consistencies were collected by three independent operators in replicates of five. The results obtained can be sumarized in
the table below:
Sample collection performance of FED
Mean Sample Weight (95% CI) 56 (55 - 57) mg
Median Sample Weight 56 mg
Range 50 ‒ 62 mg
Standard Deviation ± 3.8 mg
% Coefficient of Variation 6.9%
3. Comparing results obtained from 97 stool samples extracted by both the Fecal Extraction Device and the standard weighing
procedure. All extracts were measured with the QUANTA Flash Calprotectin Reagents (Inova Diagnostics, San Diego, USA. P/N
701350). The results obtained are summarized in tables 1 and 2, and represented in figure 1.
Table 1. Qualitative comparison between the results obtained using the Fecal Extraction Device and the standard weighing
method analizing the data using the indeterminate range as positive and negative separately.
Manual Extraction
Qualitative Comparison
Positive Indeterminate Negative Total
Positive 43 3 0 46
Fecal Extraction Indeterminate 3 20 3 26
Device Negative 0 5 20 25
Total 46 28 23 97
Indeterminate as positive
Positive agreement (95%CI): 93.2% (85.1 – 97.1%)
Negative agreement (95%CI): 87.0 % (67.9 - 95.5%)
Total agreement (95%CI): 91.8% (84.6 - 95.8%)

Indeterminate as negative
Positive agreement (95%CI): 93.5% (82.5 - 97.8%)
Negative agreement (95%CI): 94.1% (84.1 - 98.0%)
Total agreement (95%CI): 93.8% (87.2 - 97.1%)
Table 2. Quantitative comparison between the results obtained using the Fecal Extraction Device and the standard weighing
method.
Y-Intercept Slope Bias at 120mg/kg
Passing-Bablok fit Correlation r
(95% CI) (95% CI) (95% CI)
Fecal Extraction Device
-1.2 0.93 -7.8 %
vs Manual Extraction 0.975
(-9.7 to 6.4) (0.82 to 1.06) (-16.0% to 0.6%)
comparison n=97

2400
Extraction Device (mg/kg)

2000

1600

1200
Passing-Bablok fit
(y = -1.151 + 0.9321 x)
800

95% CI
400

Manual Extraction (mg/kg)

0
0 400 800 1200 1600 2000 2400
Figure 1. Linear regression analysis comparing the results obtained using the Fecal Extraction Device vs the standard weighing
procedure.

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The results obtained with the Fecal Extraction Device were equivalent to the standard weighing procedure.
The volume of the extraction buffer in the Fecal Extraction Device is 2.8 mL and, therefore, the final dilution is 1:50.

Performance Characteristics: Reproducibility

To evaluate the stool extraction reproducibility using the FED, eight samples were tested according to CLSI EP5-A3 Evaluation
of Precision of Quantitative Measurement Procedures, using three different operators. Samples were extracted every day by
each operator independently. Samples were run in replicates of 5, once a day, for 5 days, to generate 25 data points per
sample, per operator. Stool extraction reproducibility was calculated and summarized in the table 3 below.
Table 3. Fecal Extraction Device reproducibility between operators data.
Repeatability Between-Day Within Operator Between Operator Within Laboratory
Mean SD CV SD CV SD CV SD CV SD CV
Sample ID N
(mg/kg) (mg/kg) (%) (mg/kg) (%) (mg/kg) (%) (mg/kg) (%) (mg/kg) (%)
Sample 1 75 29.5 2.2 7.3% 7.2 24.4% 7.5 25.4% 0.0 0.0% 7.5 25.4%
Sample 2 75 40.0 2.3 5.7% 6.0 15.0% 6.4 16.1% 2.2 5.5% 6.8 17.0%
Sample 3 75 53.2 2.1 3.9% 5.4 10.2% 5.8 10.9% 4.8 9.0% 7.5 14.1%
Sample 4 75 108.2 4.3 4.0% 8.0 7.4% 9.1 8.4% 7.3 6.8% 11.7 10.8%
Sample 5 75 222.9 7.3 3.3% 20.9 9.4% 22.1 9.9% 5.4 2.4% 22.8 10.2%
Sample 6 75 535.4 15.7 2.9% 41.2 7.7% 44.1 8.2% 21.9 4.1% 49.2 9.2%
Sample 7 75 1278.3 45.0 3.5% 114.6 9.0% 123.2 9.6% 8.9 0.7% 123.5 9.7%
Sample 8 75 1560.0 63.9 4.1% 197.1 12.6% 207.2 13.3% 0.0 0.0% 207.2 13.3%
Greater imprecision may be observed with the FED, as compared to the predicate manual weighing method.

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Symbols Used

In Vitro diagnostic medical device Producto sanitario para diagnóstico in vitro

European Conformity Conformidad europea

Rx Only Prescription Only per US FDA Solo con prescripción, según la FDA de EE. UU

European Authorized Representative Representante europeo autorizado

Temperature Limitation Límites de temperatura

Batch Code Código de lote

Catalogue or part number Número de catálogo o componente

Manufacturer Fabricante

Use by Fecha de caducidad

Contains Sufficient Contenido suficiente para

Consult instructions for use Consultar las instrucciones de uso

∆ Indicates revision change Indica cambio de revisión

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References

1. Røseth et al.: Assessment of the neutrophil dominating protein calprotectin in feces. Scand J Gastroenterol 1992;27:793-798.

Manufactured for:

Inova Diagnostics, Inc.

9900 Old Grove Road
San Diego, CA 92131
Origin Italy
Technical Service (U.S. & Canada Only) : 877-829-4745
Technical Service (Outside the U.S.) : 1 858-805-7950

Australian Sponsor:

Werfen Australia Pty Ltd

59-61 Dickson Avenue
Artarmon NSW 2064 Australia
Tel. +61 2 9098 0200 / 1300 369 132
http://au.werfen.com/

Authorized Representative in the EU:

Representante Autorizado:

Medical Technology Promedt Consulting GmbH

Altenhofstrasse 80
66386 St. Ingbert, Germany
Tel.: +49-6894-581020
Fax.: +49-6894-581021
www.mt-procons.com

624837 April 2020

Revision 9

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