04375351190V4

ACP2
Acid Phosphatase Gen.2
• Indicates cobas c systems on which reagents can be used
Order information Roche/Hitachi cobas c systems
Acid Phosphatase Gen.2 cobas c 311 cobas c 501/502
4 x 100 tests Cat. No. 04375351 190 System-ID 07 6930 4 • •
Calibrator f.a.s. (12 x 3 mL) Cat. No. 10759350 190 Code 401
Precinorm U plus (10 x 3 mL) Cat. No. 12149435 122 Code 300
Precipath U plus (10 x 3 mL) Cat. No. 12149443 122 Code 301
Precinorm U (20 x 5 mL) Cat. No. 10171743 122 Code 300
Precipath U (20 x 5 mL) Cat. No. 10171778 122 Code 301
cobas c pack MULTI Cat. No. 04593138 190
Open/Close tool On request

English Non-prostatic acid phosphatase

R1 Connect one bottle 1 to one bottle 1a using the enclosed adapter and
System information dissolve the substrate/chromogen mixture completely in the buffer. Add a
For cobas c 311/501 analyzers: reagent tablet from bottle 2 and dissolve by gently swirling.
ACP2: ACN 021 (Total acid phosphatase)
NPP2: ACN 022 (Non-prostatic acid phosphatase) Labeling the cobas c pack MULTI
For cobas c 502 analyzer: Turn the barcode labeled side of a new cobas c pack MULTI toward you. Affix
ACP2: ACN 8021 (Total acid phosphatase) the supplied ACP2 barcode label directly over the existing barcode label.
NPP2: ACN 8022 (Non-prostatic acid phosphatase)
Intended use
In vitro test for the quantitative determination of acid phosphatase in
human serum on Roche/Hitachi cobas c systems.
Summary1,2,3
Serum acid phosphatase consists of 5 isoenzymes that originate mainly
from erythrocytes, platelets, spleen and liver reticuloendothelial cells, the
kidneys, bone, and prostate epithelial cells. Prostatic acid phosphatase
isoenzyme 2 is formed mainly, but not exclusively, in the prostate.
In general, total acid phosphatase and prostatic acid phosphatase levels
increase in the presence of progressive and metastasizing prostate carcinoma,
the increase being dependent upon the disease stage in 80 % of patients
with metastasizing prostate cancer. The percentage increase at each Filling the cobas c pack MULTI
stage depends on the classification (pathological or clinical). 1. Turn the cobas c pack MULTI toward you as shown above.
Increased acid phosphatase levels occur in Gaucher’s disease, 2. Position A of the cobas c pack is now in the center, position B on the
Niemann-Pick disease, 1-2 days after prostate surgery, biopsy, left side, position C on the right side of the cobas c pack.
manipulation or catheterization, and in the presence of benign prostate 3. Unscrew the screw cap of the bottle in position A in the middle of
hypertrophy, prostatitis and prostate infarction. the cobas c pack MULTI using the Open/Close tool.
The assay used here is a modification of the method described by Hillmann. 4. Pour the content of bottle 1 Total acid phosphatase (17 mL) into
Addition of 1,5-pentanediol increases the activity of prostatic acid phosphatase. the opened bottle of the cobas c pack (position A).
Test principle3 5. Close the bottle tightly using the Open/Close tool.
Colorimetric test 6. Unscrew the screw cap of the bottle in position B on the left side of
The 1-naphthol released during the enzymatic hydrolysis of 1-naphthyl the cobas c pack MULTI using the Open/Close tool.
phosphate is converted to an azo dye by coupling with diazotized fast red TR*. 7. Pour the content of bottle 1 Non-prostatic acid phosphatase (17 mL)
The tartrate is used as a specific inhibitor for prostatic acid phosphatase. into the opened bottle of the cobas c pack (position B).
* Fast red TR = 2-amino-5-chlorotoluene
8. Close the bottle tightly using the Open/Close tool.
acid 9. Leave position C empty.
1-naphthyl phosphate + H2O 1-naphthol + phosphate
phosphatase The ACP2 cobas c pack is now ready for use.
Note
1-naphthol + fast red TR* azo dye Use only the cobas c pack MULTI. Always use a new cobas c pack MULTI
Reagents - working solutions when preparing fresh reagent. Never reuse accessories designed for single
use, as this may result in reagent contamination and could affect test results.
R1 Bottle 1, 1a and 2:
If the cobas c pack MULTI bottles are not filled correctly, this may result
Citrate buffer: 150 mmol/L, pH 4.8; 1-naphthyl phosphate: 12.1 mmol/L;
in faulty reagent pipetting and could cause erroneous results.
fast red TR salt: 1.2 mmol/L; 1,5-pentanediol: 220 mmol/L; detergent:
3.3 mL/L; preservative Precautions and warnings
(Additionally for non-prostatic acid phosphatase determination: sodium For in vitro diagnostic use.
tartrate: 100 mmol/L) Exercise the normal precautions required for handling all laboratory reagents.
Bottle 3: Safety data sheet available for professional user on request.
Acetic acid: 0.8 mol/L Disposal of all waste material should be in accordance with local guidelines.
Reagent preparation and cobas c pack MULTI assembly
Reagent handling
Total acid phosphatase
R1 Connect one bottle 1 to one bottle 1a using the enclosed adapter and
dissolve the substrate/chromogen mixture completely in the buffer.
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ACP2
Acid Phosphatase Gen.2
Storage and stability Total acid phosphatase and Non-prostatic acid phosphatase
cobas c 501/502 test definition
ACP2
Assay type 2 Point Rate
Shelf life at 2-8 °C: See expiration date on
Reaction time / Assay points 10 / 42-70
cobas c pack label.
On-board in use and refrigerated on the analyzer: 5 days Wavelength (sub/main) 700/415 nm
Reaction direction Increase
Specimen collection and preparation Units U/L (µkat/L)
For specimen collection and preparation, only use suitable
tubes or collection containers. Reagent pipetting Diluent (H2O)
Only the specimens listed below were tested and found acceptable. R1 120 µL –
Serum. Sample volumes Sample Sample dilution
The sample types listed were tested with a selection of sample collection tubes Sample Diluent (NaCl)
that were commercially available at the time of testing, i.e. not all available Normal 10 µL – –
tubes of all manufacturers were tested. Sample collection systems from Decreased 3.3 µL – –
various manufacturers may contain differing materials which could affect Increased 20 µL – –
the test results in some cases. When processing samples in primary tubes
(sample collection systems), follow the instructions of the tube manufacturer. Calibration
Separate the serum from the clot or cells promptly. Total acid phosphatase:
Perform determinations on the samples immediately. Samples which cannot Calibrators S1: H2O
be examined immediately should be stabilized as follows: Add 1 drop S2: C.f.a.s. Use the assigned ACP2 value.
(30 µL) of solution from bottle 3 to 1.0 mL of serum and mix. Non-prostatic acid phosphatase:
Centrifuge samples containing precipitates before performing the assay. Calibrators S1: H2O
S2: C.f.a.s. Use the assigned NPP2 value.
Stability:4 8 days at 15-25 °C Calibration mode Linear
8 days at 2-8 °C 2-point calibration
Calibration frequency
4 months at (-15)-(-25) °C
- after reagent lot change
- and as required following quality control
Materials provided
procedures
See “Reagents - working solutions” section for reagents.
Traceability: This method has been standardized against the Roche system
Materials required (but not provided)
reagent using calibrated pipettes together with a manual photometer providing
See “Order information” section.
absolute values and the substrate-specific absorptivity, ε.5
General laboratory equipment
Quality control
Assay
For quality control, use control materials as listed in the
For optimum performance of the assay, follow the directions given in
“Order information” section.
this document for the analyzer concerned. Refer to the appropriate
Other suitable control material can be used in addition.
operator manual for analyzer-specific assay instructions.
The control intervals and limits should be adapted to each laboratory’s
The performance of applications not validated by Roche is not
individual requirements. Values obtained should fall within the defined
warranted and must be defined by the user.
limits. Each laboratory should establish corrective measures to be
Application for serum taken if values fall outside the limits.
Total acid phosphatase and Non-prostatic acid phosphatase Follow the applicable government regulations and local guidelines
for quality control.
cobas c 311 test definition
Assay type 2 Point Rate Calculation
Reaction time / Assay points 10 / 28-57 Roche/Hitachi cobas c systems automatically calculate the analyte
Wavelength (sub/main) 700/415 nm concentration of each sample.
Reaction direction Increase A) Total acid phosphatase: See instrument printout.
Units U/L (µkat/L) B) Prostatic acid phosphatase:
Activity Prostatic acid phosphatase =
Reagent pipetting Diluent (H2O) Activity Total acid phosphatase - Activity Non-prostatic acid phosphatase
R1 120 µL –
When measuring total acid phosphatase (ACP2) on one channel
Sample volumes Sample Sample dilution and non-prostatic acid phosphatase (NPP2) on another channel,
Sample Diluent (NaCl) the prostatic acid phosphatase can be determined directly. The
Normal 10 µL – – instrument-specific program prints out the difference between the two
Decreased 3.3 µL – – determinations as prostatic acid phosphatase.
Increased 20 µL – – Conversion factor: U/L x 0.0167 = µkat/L
Limitations - interference
Criterion: Recovery within ± 10 % of initial value at a total acid
phosphatase activity of 7 U/L (0.12 µkat/L) or at a non-prostatic acid
phosphatase activity of 4 U/L (0.07 µkat/L).
Icterus: No significant interference up to an I index of 1 (approximate
conjugated and unconjugated bilirubin concentration: 17 µmol/L (1 mg/dL)).6
Hemolysis: No significant interference up to an H index of 200 (approximate
hemoglobin concentration: 124 µmol/L (200 mg/dL)).6
Lipemia (Intralipid): No significant interference up to an L index of
200. There is poor correlation between the L index (corresponds
to turbidity) and triglycerides concentration.6
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04375351190V4

ACP2
Acid Phosphatase Gen.2
Drugs: No interference was found at therapeutic concentrations Precision
using common drug panels.7,8 Precision was determined using human samples and controls in an internal
Exception: Cefoxitine and doxycycline cause artificially high protocol. Repeatability* (n = 21), intermediate precision** (3 aliquots per run,
non-prostatic acid phosphatase results. 1 run per day, 21 days). The following results were obtained:
In very rare cases, gammopathy, in particular type IgM (Waldenström’s Total acid phosphatase:
macroglobulinemia), may cause unreliable results. Repeatability* Mean SD CV
The addition of stabilizer to the sample interferes with the U/L (µkat/L) U/L (µkat/L) %
determination of other parameters. Precinorm U 27.6 (0.461) 0.1 (0.002) 0.5
For diagnostic purposes, the results should always be assessed in conjunction Precipath U 53.1 (0.887) 0.1 (0.002) 0.2
with the patient’s medical history, clinical examination and other findings. Human serum 1 6.20 (0.104) 0.05 (0.001) 0.7
ACTION REQUIRED Human serum 2 124 (2.07) 0.4 (0.01) 0.3
Special Wash Programming: The use of special wash steps is mandatory
when certain test combinations are run together on Roche/Hitachi cobas c Intermediate Mean SD CV
systems. The latest version of the Carry over evasion list can be found with precision** U/L (µkat/L) U/L (µkat/L) %
the NaOHD/SMS/Multiclean/SCCS or the NaOHD/SMS/SmpCln1 + 2/SCCS Precinorm U 28.3 (0.473) 0.2 (0.003) 0.7
Method Sheets. For further instructions refer to the operator manual. Precipath U 53.4 (0.892) 0.5 (0.008) 0.9
cobas c 502 analyzer: All special wash programming necessary for avoiding Human serum 3 4.77 (0.080) 0.11 (0.002) 2.4
carry over is available via the cobas link, manual input is not required. Human serum 4 28.9 (0.483) 0.1 (0.002) 0.5
Where required, special wash/carry over evasion programming must
be implemented prior to reporting results with this test. Non-prostatic acid phosphatase
Limits and ranges Repeatability* Mean SD CV
Measuring range U/L (µkat/L) U/L (µkat/L) %
Total acid phosphatase and non-prostatic acid phosphatase Precinorm U 13.1 (0.219) 0.1 (0.002) 0.7
0.5-200 U/L (0.01-3.34 µkat/L) Precipath U 35.2 (0.588) 0.1 (0.002) 0.4
Determine samples having higher activities via the rerun function. Dilution of Human serum 1 3.18 (0.053) 0.04 (0.007) 1.3
samples via the rerun function is a 1:3 dilution. Results from samples diluted Human serum 2 13.7 (0.229) 0.1 (0.002) 0.5
by the rerun function are automatically multiplied by a factor of 3.
Intermediate Mean SD CV
Lower limits of measurement
precision** U/L (µkat/L) U/L (µkat/L) %
Lower detection limit of the test
0.5 U/L (0.01 µkat/L) Precinorm U 13.4 (0.224) 0.1 (0.002) 1.0
The lower detection limit represents the lowest measurable analyte Precipath U 35.1 (0.586) 0.4 (0.007) 1.1
level that can be distinguished from zero. It is calculated as the value Human serum 3 3.00 (0.050) 0.1 (0.002) 4.6
lying three standard deviations above that of the lowest standard Human serum 4 18.5 (0.309) 0.2 (0.003) 0.8
(standard 1 + 3 SD, repeatability, n = 21). * repeatability = within-run precision
Expected values ** intermediate precision = total precision / between run precision / between day precision
Total acid phosphatase (37 °C)9 Method comparison
Men < 6.6 U/L (< 0.110 µkat/L) Total acid phosphatase and non-prostatic acid phosphatase
Women < 6.5 U/L (< 0.108 µkat/L) Acid phosphatase values for human serum samples obtained on a
Prostatic acid phosphatase (37 °C)9 Roche/Hitachi cobas c 501 analyzer (y) were compared with those determined
Men < 3.5 U/L (< 0.058 µkat/L) using the same reagent on a Roche/Hitachi 917 analyzer (x).
Each laboratory should investigate the transferability of the expected values to Total acid phosphatase:
its own patient population and if necessary determine its own reference ranges. Sample size (n) = 66
Passing/Bablok10 Linear regression
Specific performance data
Representative performance data on the analyzers are given below. y = 0.999x + 0.045 U/L y = 0.977x + 0.766 U/L
Results obtained in individual laboratories may differ. τ = 0.994 r = 1.000
The sample activities were between 4.38 and 190 U/L (0.073 and 3.17 µkat/L).
Non-prostatic acid phosphatase:
Sample size (n) = 72
Passing/Bablok10 Linear regression
y = 0.971x - 0.010 U/L y = 0.957x + 0.292 U/L
τ = 0.980 r = 0.999
The sample activities were between 2.32 and 161 U/L (0.039 and 2.69 µkat/L).
References
1. Tietz NW. Clinical Guide to Laboratory Tests, 3rd ed. Philadelphia,
Pa: WB Saunders Company, 1995:516-519.
2. Heller JE. Prostatic acid phosphatase: Its current clinical
status. J Urol 1987;137:1091-1103.
3. Hillmann G. Continuous Photometric Measurement of Prostate
Phosphatase Activity. Zeitschrift fuer Klinische Chemie und Klinische
Biochemie, 1971, vol. 9, no. 3, p. 273-274.
4. Guder WG, Narayanan S, Wisser H, Zawta B. List of Analytes;
Preanalytical Variables. Brochure in: Samples: From the Patient
to the Laboratory. Darmstadt: GIT-Verlag, 1996.
5. Data on file at Roche Diagnostics.
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ACP2
Acid Phosphatase Gen.2
6. Glick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences
in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
7. Breuer J. Report on the Symposium “Drug effects in Clinical Chemistry
Methods”. Eur J Clin Chem Clin Biochem 1996;34:385-386.
8. Sonntag O, Scholer A. Drug interference in clinical chemistry:
recommendation of drugs and their concentrations to be used in drug
interference studies. Ann Clin Biochem 2001;38:376-385.
9. Junge W, Thormeyer I, Schlottmann A et al. Determination of
Reference Values for Acid Phosphatase using a New Photometric
Assay. Pecs, Hungary: 3rd Alpe-Adria Congress on Clinical Chemistry
and Laboratory Medicine. September 7-9, 1994.
10. Passing H, Bablok W et al. A General Regression Procedure for Method
Transformation. J Clin Chem Clin Biochem 1988;26:783-790.

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cobas c systems 4/4 2010-05, V 4 English