BIOMÉRIEUX

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®
VIDAS TPSA (TPSA)

INTENDED USE
VIDAS® TPSA is an automated quantitative test for use on the VIDAS® family instruments, for the quantitative
measurement of prostate specific antigen (PSA) levels in human serum or plasma (lithium heparin or EDTA), using the
ELFA technique (Enzyme Linked Fluorescent Assay).
SUMMARY AND EXPLANATION
Prostate-specific antigen (PSA) is a glycoprotein which belongs to the kallikrein family.1,2,3 PSA has a molecular weight
of 30,000 daltons.3,4,5
PSA is principally produced by the glandular epithelium of the prostate, and is secreted in the seminal fluid. PSA is also
present in urine and blood. PSA acts on seminal fluid to fluidify and increase sperm mobility.2,3,6
PSA levels rise in prostatic pathologies such as benign prostatic hyperplasia (BPH) or prostate cancer. Testing for PSA
and its evolution is useful for monitoring and controlling the efficacy of prostatic carcinoma therapy.5,6 Determination of
PSA levels enables the detection of the onset of metastases or the persistence of disease following prostate cancer
therapy. An elevated PSA level after therapy or a persistently high level during therapy indicates residual or recurrent
disease.
PSA is present in blood with three main forms. The most important immunoreactive form is PSA bound to
alpha‑1‑antichymotrypsin (PSA-ACT). Free PSA is the other immunoreactive form present in serum. Equimolar PSA
assays detect the bound form (PSA-ACT) and the free form in the same manner.4,5 The VIDAS® TPSA assay is an
equimolar test7: the molar ratio (concentration of solution containing 100% Free PSA over concentration of solution
containing 100% PSA-ACT) is between 105 and 125%.
The third form of PSA, bound to alpha-2-macroglobulin, cannot be detected by immunoassays.
The VIDAS® TPSA assay is used in the diagnosis of prostate disorders, including cancer of the prostate, and for the
prognosis and monitoring of patients with diagnosed malignant tumors.
PRINCIPLE
The assay principle combines a two-step sandwich enzyme immunoassay method with a final fluorescence detection
(ELFA).
The single-use Solid Phase Receptacle (SPR) serves as the solid phase as well as the pipetting device. Reagents for the
assay are ready-to-use and pre-dispensed in the sealed single-use reagent strips.
All of the assay steps are performed automatically by the instrument. The reaction medium is cycled in and out of the
SPR device several times.
The sample/conjugate mixture is cycled in and out of the SPR device several times.
This operation enables the antibody fixed onto the interior wall of the SPR device to capture the prostate specific antigen
present in the sample. Unbound components are eliminated during washing steps. Alkaline phosphatase-labeled
antibody is then incubated in the SPR device where it binds with the prostate specific antigen. Unbound conjugate is then
eliminated during the washing steps.
During the final detection step, the substrate (4-Methyl-umbelliferyl phosphate) is cycled in and out of the SPR device.
The conjugate enzyme catalyzes the hydrolysis of this substrate into a fluorescent product (4-Methyl-umbelliferone), the
fluorescence of which is measured at 450 nm. The intensity of the fluorescence is proportional to the concentration of
prostate specific antigen present in the sample.
At the end of the assay, the results are automatically calculated by the instrument according to the calibration curve
stored in memory. The results can then be printed out.
CONTENT OF THE KIT (60 TESTS) - RECONSTITUTION OF REAGENTS

60 TPSA Strips(a) STR Ready-to-use.

60 TPSA Solid Phase SPR Ready-to-use.
Receptacles Interior of SPR device coated with monoclonal anti-PSA immunoglobulins
2 x 30 (mouse).

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TPSA Calibrator(b) S1 Reconstitute with 2 mL of distilled water. Let stand for 30 minutes, then mix.
1 x 2 mL After reconstitution, stable for 24 hours at +2°C/+8°C or until the expiration
(lyophilized) date on the kit at -25°C ± 6°C. 5 freeze/thaw cycles are possible.
Human serum* + human PSA + preservatives.
MLE data indicate the concentration in ng/mL (“Calibrator (S1) Dose Value”)
and the confidence interval in “Relative Fluorescence Value” (“Calibrator (S1)
RFV Range”).
TPSA Control(b) C1 Reconstitute with 2 mL of distilled water. Let stand for 30 minutes, then mix.
1 x 2 mL After reconstitution, stable for 24 hours at +2°C/+8°C or until the expiration
(lyophilized) date on the kit at -25°C ± 6°C. 5 freeze/thaw cycles are possible.
Human serum* + human PSA + preservatives.
MLE data indicate the confidence interval in ng/mL ("Control C1 Dose Value
Range").
TPSA Sample Diluent R1 Ready-to-use.
2 x 4 mL Calf serum + 0.9 g/L sodium azide.

(liquid)
Specifications for the factory master data required to calibrate the assay: MLE (Master Lot Entry) barcode printed on
the box label.
1 package insert downloadable from www.biomerieux.com/techlib

* This product has been tested and shown to be negative for HBs antigen, antibodies to HIV1, HIV2 and HCV. However,
since no existing test method can totally guarantee their absence, this product must be treated as potentially infectious.
Therefore, usual safety procedures should be observed when handling.

(a) DANGER H318 / P280 / P305 + P351 + P338

(b) WARNING EUH208 / H317 / P261 / P280 / P302 + P352

Hazard statements
• EUH208: Contains 2-methyl-2H-isothiazolin-3-one. May produce an allergic reaction.
• H317: May cause an allergic skin reaction.
• H318: Causes serious eye damage.

Precautionary statements
• P261: Avoid breathing dust/fume/gas/mist/vapours/spray.
• P280: Wear protective gloves/protective clothing/eye protection/face protection.
• P302 + P352: IF ON SKIN: Wash with plenty of soap and water.
• P305 + P351 + P338: IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if
present and easy to do. Continue rinsing.

For further information, consult the Safety Data Sheet.

The SPR device

The SPR device is coated during production with monoclonal anti-PSA antibodies (mouse). Each SPR device is
identified by the TPSA code. Only remove the required number of SPR devices from the pouch and carefully reseal the
pouch after opening.

The Reagent Strip

The strip consists of 10 wells covered with a labeled foil seal. The label comprises a barcode which mainly indicates the
assay code, kit lot number, and expiration date. The foil of the first well is perforated to facilitate the introduction of the

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sample. The last well of each strip is a cuvette in which the fluorometric reading is performed. The wells in the center
section of the strip contain the various reagents required for the assay.

Description of the TPSA strip

Well Reagents
1 Sample well.
2-3-4-9 Empty wells.
5 Conjugate: Alkaline phosphatase-labeled monoclonal anti-PSA immunoglobulins (mouse) +
0.9 g/L sodium azide (400 µL).
6-7 Wash buffer: Tris (0.05 mol/L, pH 7.4) + NaCl (0.4 mol/L) + Tween (0.05%) + 0.9 g/L sodium
azide (600 µL).
8 Diluent: Tris (0.1 mol/L) + NaCl (0.1 mol/L) + calf serum (5%) + 0.9 g/L sodium azide (400 µL).
10 Reading cuvette with substrate: 4‑Methyl‑umbelliferyl phosphate (0.6 mmol/L) + diethanolamine
(DEA) (0.62 mol/L or 6.6%, pH 9.2) + 1 g/L sodium azide (300 μL).

MATERIALS AND DISPOSABLES REQUIRED BUT NOT PROVIDED

• Pipette with disposable tip to dispense 2 mL and 200 µL.
• Powderless disposable gloves.
• For other specific materials and disposables, please refer to the Instrument User Manual.
• Instruments of the VIDAS® family.

WARNINGS AND PRECAUTIONS

• For in vitro diagnostic use only.
• For professional use only.
• This kit contains products of human origin. No known analysis method can totally guarantee the absence of
transmissible pathogenic agents. It is therefore recommended that these products be treated as potentially
infectious and handled observing the usual safety precautions (see Laboratory Biosafety Manual - WHO -
Geneva - latest edition).
• This kit contains products of animal origin. Certified knowledge of the origin and/or sanitary state of the animals does
not totally guarantee the absence of transmissible pathogenic agents. It is therefore recommended that these
products be treated as potentially infectious, and handled observing the usual safety precautions (do not ingest; do
not inhale).
• Do not use the SPR devices if the pouch is pierced or if the dot sealing a SPR device has come unstuck.
• Do not use visibly deteriorated strips (damaged foil or plastic).
• Do not use reagents after the expiration date indicated on the box label.
• Do not mix reagents (or disposables) from different lots.
• Use powderless gloves, as powder has been reported to cause false results for certain enzyme immunoassay tests.
• Kit reagents contain sodium azide which can react with lead or copper plumbing to form explosive metal azides. If
any liquid containing sodium azide is disposed of in the plumbing system, drains should be flushed with water to
avoid build-up.
• Refer to the hazard statements “H” and precautionary statements “P” indicated above.
• Spills should be wiped up thoroughly after treatment with liquid detergent or a solution of household bleach
containing at least 0.5% sodium hypochlorite. See the User Manual for cleaning spills on or in the instrument. Do not
autoclave solutions containing bleach.
• The instrument should be regularly cleaned and decontaminated (refer to the User Manual for user and preventive
maintenance operations).

STORAGE CONDITIONS
• Store the kit at +2°C/+8°C.
• Do not freeze SPR devices and strips.
• Store all unused reagents at +2°C/+8°C.
• After opening the kit, check that the SPR pouch is correctly sealed and undamaged. If not, do not use the SPR
devices.
• After use, carefully reseal the pouch with the desiccant inside to maintain stability of the SPR devices, and
return the complete kit to +2°C/+8°C.
• If stored according to the recommended conditions, all components are stable until the expiration date indicated on
the box label. Refer to the kit composition table for special storage conditions.

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SAMPLES
Specimen type and collection
Human serum or plasma (lithium heparin or EDTA)
As some collection tubes contain substances which interfere with test results, it is recommended that each laboratory
checks the compatibility of collection tubes used.
Samples containing impurities must be centrifuged before analysis.
None of the following factors have been found to significantly influence this assay:
• hemolysis (after spiking samples with hemoglobin: 0 to 300 µmol/L (monomer)),
• lipemia (after spiking samples with lipids: 0 to 10 mg/mL equivalent in triglycerides),
• bilirubinemia (after spiking samples with bilirubin: 0 to 500 µmol/L).

However, it is recommended not to use samples that are clearly hemolyzed, lipemic or icteric, and if possible to collect a
new sample.
Sample stability
Samples can be stored at +2°C/+8°C in stoppered tubes for a maximum of 24 hours; if longer storage is required, freeze
the sera or plasma at -25°C ± 6°C. A study performed on frozen samples over a period of 2 months showed that the
quality of results is not affected. Avoid successive freezing and thawing.
INSTRUCTIONS FOR USE
For complete instructions, see the Instrument User Manual.
Reading VIDAS® PTC (Protocol Test Change) data and MLE data

When using the assay for the first time

With the external instrument barcode reader, scan the barcodes (PTC and MLE) in the following order:
1. According to the instrument used, scan the PTC barcode(s) downloadable from www.biomerieux.com/techlib. This
reading allows VIDAS® PTC protocol data to be transferred to the instrument software for its update.
2. Scan the MLE data on the box label.

When opening a new lot of reagents

With the external instrument barcode reader, scan the MLE data on the box label before performing the test.
Note: The master lot data need only be entered once for each lot.
It is possible to enter MLE data manually or automatically depending on the instrument (refer to the User Manual).
Calibration
Calibration, using the calibrator provided in the kit, must be performed each time a new lot of reagents is opened, after
the MLE data have been entered, and then every 56 days. This operation provides instrument-specific calibration curves
and compensates for possible minor variations in assay signal throughout the shelf life of the kit.
The calibrator, identified by S1, must be tested in duplicate. The calibrator value must be within the set RFV (Relative
Fluorescence Value) range. If this is not the case, recalibrate.
VIDAS® TPSA is calibrated against the reference standard: 1st WHO international standard 96/670.8 Depending on the
dilution mode and the type of diluent used with the international standard, a bias of up to 15% may be observed.
Procedure
1. Only remove the required reagents from the refrigerator and allow them to come to room temperature for at
least 30 minutes.
2. Use one "TPSA" strip and one "TPSA" SPR device for each sample, control or calibrator to be tested. Make sure the
SPR pouch has been carefully resealed after the required SPR devices have been removed.
3. The test is identified by the "TPSA" code on the instrument. The calibrator, identified by S1, must be tested in
duplicate. If the control needs to be tested, it should be identified by C1.
4. Mix the calibrator, control and samples using a vortex-type mixer (for serum or plasma separated from the pellet).
5. For this test, the calibrator, control and sample test portion is 200 µL.
6. Insert the "TPSA" SPR devices and "TPSA" strips into the instrument. Check to make sure the color labels with the
assay code on the SPR devices and the Reagent Strips match.
7. Initiate the assay as directed in the User Manual. All the assay steps are performed automatically by the instrument.
8. Close the vials and return them to the required temperature after pipetting.

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9. The assay will be completed within approximately 60 minutes. After the assay is completed, remove the SPR
devices and strips from the instrument.
10. Dispose of the used SPR devices and strips into an appropriate recipient.
RESULTS AND INTERPRETATION
Once the assay is completed, results are analyzed automatically by the computer. Fluorescence is measured twice in the
Reagent Strip’s reading cuvette for each sample tested. The first reading is a background reading of the substrate
cuvette before the SPR device is introduced into the substrate.
The second reading is taken after incubating the substrate with the enzyme bound to the interior of the SPR device.
The RFV (Relative Fluorescence Value) is calculated by subtracting the background reading from the final result. This
calculation appears on the result sheet.
The results are automatically calculated by the instrument using calibration curves which are stored by the instrument
(4‑parameter logistic model); the concentrations are expressed in ng/mL.
Samples with TPSA test value concentrations > 100 ng/mL should be reassayed after diluting with TPSA sample diluent
(R1).
If the dilution factor has not been entered when the Work List was created (see User Manual), multiply the result by the
dilution factor to obtain the sample concentration.
Interpretation of test results should be made taking into consideration the patient's clinical history, and the results of any
other tests performed.
QUALITY CONTROL
One control is included in this kit.
This control must be performed immediately after opening a new kit to ensure that reagent performance has not been
altered. Each calibration must also be checked using this control. The instrument will only be able to check the control
value if it is identified by C1. Results cannot be validated if the control value deviates from the expected values.
Note: It is the responsibility of the user to perform Quality Control in accordance with any local applicable regulations.
LIMITATIONS OF THE METHOD
Interference may be encountered with certain sera containing antibodies directed against reagent components. For this
reason, assay results should be interpreted as part of a complete clinical profile.
The serum PSA value in an isolated specimen can only be used in conjunction with clinical data and information
available from other diagnostic procedures. An abnormal PSA level does not necessarily signify a malignant disorder.
TPSA assays should not be performed on patients who have received a contrast medium within the previous 24 hours.9
RANGE OF EXPECTED VALUES
Expected values were determined using 1041 samples collected from healthy subjects. PSA concentrations per age
group are as follows:
PSA concentrations (ng/mL)
Age (years) Low limit* High limit*
< 40 0.21 1.72
40 - 49 0.27 2.19
50 - 59 0.27 3.42
60 - 69 0.22 6.16
> 69 0.21 6.77

* including 95% of the population.

For a healthy male population aged 50 years or more, the distribution of values is as follows:
PSA (ng/mL) 0-2 2-4 4-6 >6
Frequency 83.5% 13.5% 1.3% 1.7%

For a population of 54 patients with benign prostate hypertrophy, the distribution of values is as follows:

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Distribution in percentage (%) in relation to the zone of

No. of subjects values in ng/mL
< 4 ng/mL 4-10 ng/mL > 10 ng/mL
Mean = 3.08 ng/mL 74 24 2
54
Standard deviation = 2.3 ng/mL (n=40) (n=13) (n=1)

These results are given as a guide; it is recommended that each laboratory establish its own reference values from a
rigorously selected population.
PERFORMANCE
Measurement range
The measurement values of the VIDAS® TPSA kit range from 0.07 to 100 ng/mL.
Detection limit
Defined as the smallest concentration of prostate specific antigen which is significantly different from the zero
concentration with a probability of 95%: 0.07 ng/mL.
Hook effect
No hook effect was found up to prostate specific antigen concentrations of 100,000 ng/mL.
Precision
The results presented in the following tables are given for information purposes only.
Within-Run Reproducibility (intra-assay)
Five samples were tested 30 times in the same run.
Sample 1 2 3 4 5
Mean (ng/mL) 0.55 3.0 7.6 17.4 31.6
CV (%) 6.5 3.9 3.5 3.4 3.9

Between-Run Reproducibility (inter-assay)

Five samples were tested in 29 different runs on the same VIDAS® instrument.
Sample 1 2 3 4 5
Mean (ng/mL) 0.52 2.8 7.5 16.9 32.1
CV (%) 6.8 3.9 3.9 4.3 5.5

Accuracy
Dilution test
The serum matrix of the sample can influence the results of the dilution test. When printing out results, it is
recommended to indicate the level of dilution used.
Three samples were diluted in the TPSA diluent and tested singly in 3 runs. The measured mean concentration
compared to the expected mean concentration is expressed as a mean recovery percentage.
Sample Dilution factor Expected mean Measured mean Mean recovery
concentration concentration percentage
(ng/mL) (ng/mL) (%)
1 1/1 92.9 92.9 100
1/2 46.4 41.9 90
1/4 23.2 19.8 85
1/8 11.6 9.9 85
1/16 5.8 5.0 86

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Sample Dilution factor Expected mean Measured mean Mean recovery

concentration concentration percentage
(ng/mL) (ng/mL) (%)
2 1/1 52.8 52.8 100
1/2 26.4 26.1 99
1/4 13.2 12.5 95
1/8 6.6 6.4 97
1/16 3.3 3.2 97
3 1/1 18.2 18.2 100
1/2 9.1 8.1 89
1/4 4.6 4.2 92
1/8 2.3 2.2 96
1/16 1.1 1.1 96

Diagnostic Specificity and Sensitivity

109 samples from patients with benign prostatic hyperplasia and 205 from patients with prostate cancer were tested
using VIDAS® TPSA.
A ROC curve was established using the 314 samples. 2 cut-off values were identified:
• 3.03 ng/mL: sensitivity = 95%
• 6.73 ng/mL: sensitivity = 80%

Cut-Off Value

3.03 ng/mL 6.73 ng/mL

[95%CI]* [95%CI]
Sensitivity 91.16 97.36 73.86 84.99
Specificity 9.88 23.75 46.41 65.09
Positive Predictive Value (PPV) 67.94 77.36
Negative Predictive Value (NPV) 60.71 59.22

* CI: Confidence Interval

Comparison with Other Test Methods
The concentration of prostate specific antigen may vary in a sample determined using kits from different manufacturers,
depending on the test methods used.
If the test method is changed, and in the case of patient monitoring, laboratories should confirm the concentrations
previously found.
Correlation was established between VIDAS® TPSA (Y), and another enzyme immunoassay method (X):
X = 0,99 Y - 0,17 r = 0,994 (n = 126)
WASTE DISPOSAL
Dispose of used or unused reagents, as well as any other contaminated disposable materials, following procedures for
infectious or potentially infectious products.
It is the responsibility of each laboratory to handle waste and effluents produced, according to their nature and degree of
hazardousness, and to treat and dispose of them (or have them treated and disposed of) in accordance with any
applicable regulations.
LITERATURE REFERENCES
1. KANTOFF P.W. and TALCOT J.A. The prostate specific antigen. Its use as a tumor marker for prostate cancer.
Hematol Oncol Clin N Amer 1994; 8: 555-72.
2. OSTERLING JE. Prostate specific antigen: A critical assessment of the most useful tumor marker for
adenocarcinoma of the prostate. J Urol 1991; 145: 907-23.

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3. CHRISTENSON A., LAURELL C-B., LILIJA H. Enzymatic activity of Prostate-specific Antigen and its reactions with
extracellular Serine Proteinase inhibitor. Eur J Biochem 1990; 194: 755-63.
4. ZHANG W.M., LEINONEN J., KALKKINEN N., et al. Purification and Characterization of different Molecular forms of
Prostate-specific antigen in human Seminal fluid. Clin Chem 1995; 41/11: 1567-1573
5. STENMAN U-H., LEINONEN J., ZHANG W-H., Problems in the determination of Prostate specific antigen. Eur J Clin
Chem Clin Biochem 1996; 34: 735-40.
6. STAMEY TA. YANG N., HAY AR. et al. Prostate specific antigen as a serum marker for adenocarcinoma of the
prostate. New Engl J Med 1987, 317: 909-16.
7. STAMEY T. A., PRESTIGIACOMO A. F. and CHEN Z. Standardization of immunoassays for prostate specific antigen,
a different view based on experimental observations. Cancer, 1994, 74, 1662-1666.
8. RAFFERTY B., RIGSBY P., ROSE. M., et al. Reference Reagents for Prostate-specific antigen (PSA): Establishment
of the First International Standards for free PSA and PSA (90: 10). Clinical Chemistry, 2000, vol. 46, 1310-1317.
9. WATANABE N. et al. In vitro effect of contrast during immunoradiometric assay for tumour-associated antigens.
Nuclear Medicine Communication, 1998, 19, 63-70.
INDEX OF SYMBOLS

Symbol Meaning
Catalogue number

In Vitro Diagnostic Medical Device

Manufacturer

Temperature limit

Use by date

Batch code

Consult Instructions for Use

Contains sufficient for <n> tests

Date of manufacture

LIMITED WARRANTY
bioMérieux warrants the performance of the product for its stated intended use provided that all procedures for usage,
storage and handling, shelf life (when applicable), and precautions are strictly followed as detailed in the instructions for
use (IFU).
Except as expressly set forth above, bioMérieux hereby disclaims all warranties, including any implied warranties of
merchantability and fitness for a particular purpose or use, and disclaims all liability, whether direct, indirect or
consequential, for any use of the reagent, software, instrument and disposables (the "System") other than as set forth in
the IFU.
REVISION HISTORY
Change type categories
N/A Not applicable (First publication)
Correction Correction of documentation anomalies
Technical change Addition, revision and/or removal of information related to the product
Administrative Implementation of non-technical changes noticeable to the user

Note: Minor typographical, grammar, and formatting changes are not included in the revision
history.

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Release Date Part Number Change Type Change Summary

Administrative LIMITED WARRANTY
2018/02 09296J
Technical change INSTRUCTIONS FOR USE
CONTENT OF THE KIT (60 TESTS)
2019-10 050883-02 Technical change
WARNINGS AND PRECAUTIONS
Administrative Formatting and wording changes.
CONTENT OF THE KIT (60 TESTS)
2020-08 050883-03 CALIBRATION
Technical change
PERFORMANCE

BIOMERIEUX, the BIOMERIEUX logo, SPR and VIDAS are used, pending, and/or registered trademarks belonging to
bioMérieux, or one of its subsidiaries or one of its companies.
Any other name or trademark is the property of its respective owner.

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