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LACTATE DEHYDROGENASE

7D69-20
30-3133/R4

LACTATE
DEHYDROGENASE
This package insert contains information to run the Lactate Dehydrogenase assay on the ARCHITECT c Systems™
and the AEROSET System.

NOTE: Changes Highlighted

NOTE: This package insert must be read carefully prior to product use. Package insert instructions must be followed
accordingly. Reliability of assay results cannot be guaranteed if there are any deviations from the instructions in
this package insert.

Customer Support
United States: 1-877-4ABBOTT
Canada: 1-800-387-8378 (English speaking customers)
1-800-465-2675 (French speaking customers)
International: Call your local Abbott representative

Symbols in Product Labeling

Concentration Catalog number/List number


Authorized Representative in the
Serial number
European Community
Ingredients Consult instructions for use

In vitro diagnostic medical device Manufacturer

Batch code/Lot number Temperature limitation

Reagent 1 Use by/Expiration date

Reagent 2

ABBOTT LABORATORIES ABBOTT


Abbott Park, IL 60064, USA Max-Planck-Ring 2
65205 Wiesbaden
Germany
+49-6122-580
October 2006
©2002, 2006 Abbott Laboratories

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NAME WARNINGS AND PRECAUTIONS
LACTATE DEHYDROGENASE Precautions for Users
1. For in vitro diagnostic use.
INTENDED USE
2. Hemolyzed specimens are not recommended for use.
The Lactate Dehydrogenase (LD) assay is used for the quantitation of
lactate dehydrogenase in human serum or plasma. 3. Do not use components beyond the expiration date.
4. Do not mix materials from different kit lot numbers.
SUMMARY AND EXPLANATION OF TEST 5. CAUTION: This product requires the handling of human specimens.
Lactate dehydrogenase (LD) is an enzyme which can be found in most It is recommended that all human sourced materials be considered
major tissues. Serum levels of LD are elevated in a wide variety of potentially infectious and handled in accordance with the OSHA
pathologic conditions, most notably cardiac and hepatic disease. LD is Standard on Bloodborne Pathogens.3 Biosafety Level 24 or other
a tetrameric enzyme consisting of two basic subunits. Five isoenzymes appropriate biosafety practices5,6 should be used for materials that
can be observed after electrophoresis. The relative ratios of the contain or are suspected of containing infectious agents
isoenzymes vary with the tissue source of the LD.1 NOTE: Refer to Section 8 of the instrument-specific operations manual
for proper handling and disposal of reagents containing sodium azide.
PRINCIPLES OF PROCEDURE For product not classified as dangerous per European Directive
LD catalyzes the conversion of lactate to pyruvate, the forward reaction 1999/45/EC as amended, safety data sheet available for professional
and the conversion of pyruvate to lactate, the reverse reaction. The user on request.
enzyme may be assayed using either material as a substrate; however,
the enzyme activities obtained by the two methods are not directly SPECIMEN COLLECTION AND HANDLING
comparable. The forward reaction does not require preincubation to
exhaust endogenous α-keto acids and displays linearity over a wider Suitable Specimens
range of activity in patient samples. This LD method utilizes the forward Serum and plasma are acceptable specimens.
reaction.2 • Serum: Use serum collected by standard venipuncture techniques
Lactate and NAD+ are converted to pyruvate and NADH by the action into glass or plastic tubes with or without gel barriers. Ensure
of LD. NADH strongly absorbs light at 340 nm, whereas NAD+ does not. complete clot formation has taken place prior to centrifugation.
The rate of increase in absorbance at 340 nm is directly proportional to Separate serum from red blood cells or gel as soon after collection
the LD activity in the sample. as possible. Hemolyzed serum must not be used because
Methodology: Lactate to Pyruvate (NADH) erythrocytes contain 150 times more LD activity than serum.1
Some specimens, especially those from patients receiving
REAGENTS anticoagulant or thrombolytic therapy, may take longer to complete
their clotting processes. Fibrin clots may subsequently form in these
Reagent Kit sera and the clots could cause erroneous test results.
7D69 Lactate Dehydrogenase is supplied as a liquid, ready-to-use,
• Plasma: Use plasma collected by standard venipuncture techniques
two-reagent kit which contains:
into glass or plastic tubes without gel barriers. Acceptable
10 x 70 mL anticoagulants are lithium heparin (with or without gel barrier)
10 x 21 mL and sodium heparin. Ensure centrifugation is adequate to remove
platelets. Separate plasma from red blood cells or gel as soon after
Estimated tests per kit: 3,621 collection as possible. Hemolyzed plasma must not be used because
Calculation is based on the minimum reagent fill volume per kit. erythrocytes contain 150 times more LD activity than serum.1
Reactive Ingredients Concentration Refer to the specimen collection tube manufacturer’s instructions for
processing and handling requirements.
Diethanolamine 381 mmol/L
L-Lactate, Lithium Salt 76 mmol/L For total sample volume requirements, refer to the instrument-specific
ASSAY PARAMETERS section of this package insert and Section 5 of
Sodium Azide < 0.1% the instrument-specific operations manual.
β-NAD 30.8 mmol/L Specimen Storage
Serum and plasma
REAGENT HANDLING AND STORAGE
Reagent Handling Temperature Maximum Bibliographic
Remove air bubbles, if present in the reagent cartridge, with a new Storage Reference
applicator stick. Alternatively, allow the reagent to sit at the appropriate 20 to 25°C 7 days 7
storage temperature to allow the bubbles to dissipate. To minimize
volume depletion, do not use a transfer pipette to remove the bubbles. 2 to 8°C 4 days 7, 8
CAUTION: Reagent bubbles may interfere with proper detection of -20°C 6 weeks 7
reagent level in the cartridge, causing insufficient reagent aspiration
which could impact results. Guder et al.7
suggest storage of frozen specimens at -20°C for
no longer than the time interval cited above. However, limitations
Reagent Storage of laboratory equipment make it necessary in practice for clinical
Unopened reagents are stable until the expiration date when stored at laboratories to establish a range around -20°C for specimen storage.
2 to 8°C. This temperature range may be established from either the freezer
manufacturer’s specifications or your laboratory standard operating
Reagent stability is 30 days if the reagent is uncapped and onboard.
procedure(s) for specimen storage.
NOTE: Stored specimens must be inspected for particulates. If present,
mix and centrifuge the specimen to remove particulates prior to testing.

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PROCEDURE RESULTS
Materials Provided Refer to the instrument-specific operations manual for information on
results calculations.
7D69 Lactate Dehydrogenase Reagent Kit
• ARCHITECT System Operations Manual—Appendix C
Materials Required but not Provided • AEROSET System Operations Manual—Appendix A
• Control Material Representative performance data are given in the EXPECTED VALUES
• Saline (0.85% to 0.90% NaCl) for specimens that require dilution and SPECIFIC PERFORMANCE CHARACTERISTICS sections of this
Assay Procedure package insert. Results obtained in individual laboratories may vary.
For a detailed description of how to run an assay, refer to Section 5 of LIMITATIONS OF THE PROCEDURE
the instrument-specific operations manual.
Refer to the SPECIMEN COLLECTION AND HANDLING and SPECIFIC
Specimen Dilution Procedures PERFORMANCE CHARACTERISTICS sections of this package insert.
The ARCHITECT c Systems and the AEROSET System have automatic
dilution features; refer to Section 2 of the instrument-specific operations EXPECTED VALUES
manual for additional information.
Reference Range
Serum and plasma: Specimens with LD values exceeding 1,984 U/L
(4,732 U/L for Flex Rate Linearity) are flagged and may be diluted using Serum/Plasma9
the Automated Dilution Protocol or the Manual Dilution Procedure. Range (U/L)
Automated Dilution Protocol Adult 125 to 243
If using the Automated Dilution Protocol, the system performs a 1:5 A study was conducted using 124 serum samples and 122 plasma
dilution of the specimen and automatically corrects the enzyme activity samples drawn from 74 female and 50 male volunteers. Data were
value by multiplying the result by the appropriate dilution factor. analyzed as described by Clinical and Laboratory Standards Institute
Manual Dilution Procedure (CLSI) protocol NCCLS C28-A.10 From this study, 95% of specimens fell
within 125 to 243 U/L, with samples ranging from 115 to 287 U/L.
Manual dilutions should be performed as follows:
It is recommended that each laboratory determine its own reference
• Use saline (0.85% to 0.90% NaCl) to dilute the sample. range based upon its particular locale and population characteristics.
• The operator must enter the dilution factor in the patient or control
order screen. The system uses this dilution factor to automatically SPECIFIC PERFORMANCE CHARACTERISTICS
correct the enzyme activity value by multiplying the result by the
entered factor. Linearity
• If the operator does not enter the dilution factor, the result must be Lactate Dehydrogenase is linear up to 1,984 U/L.
multiplied by the appropriate dilution factor before reporting the result. Flex Rate Linearity is 4,732 U/L. To use Flex Rate Linearity, the operator
NOTE: If a diluted sample result is flagged indicating it is less than the must edit the linear high value to 4,732 on the appropriate screen.
linear low limit, do not report the result. Rerun using an appropriate • ARCHITECT c Systems—Configure assay parameters screen,
dilution. Results view
For detailed information on ordering dilutions, refer to Section 5 of the • AEROSET System—Assay Configuration screen, Outline page
instrument-specific operations manual. Linearity was verified using CLSI protocol NCCLS EP6-P.11

CALIBRATION Limit of Detection (LOD)


Calibration is stable for approximately 30 days (720 hours) and is The LOD for Lactate Dehydrogenase is 5.0 U/L. The LOD is the mean
required with each change in reagent lot number. Verify calibration with concentration of an analyte-free sample + 2 SD, where SD = the
at least two levels of controls according to the established quality control pooled, within-run standard deviation of the analyte-free sample. A
requirements for your laboratory. If control results fall outside acceptable study performed on an ARCHITECT c System and an AEROSET System
ranges, recalibration may be necessary. produced an LOD for Lactate Dehydrogenase of 3.1 U/L.
A calibration factor must be entered. Refer to the PARAMETERS Limit of Quantitation (LOQ)
section for the specific factor. The LOQ for Lactate Dehydrogenase serum is 7.7 U/L. The LOQ is the
• ARCHITECT c Systems—Configure assay parameters window, analyte concentration at which the CV = 20%.
Calibration view
Interfering Substances12
• AEROSET System—Assay Configuration screen, Calibration page
Interference studies were conducted using CLSI protocol NCCLS
For a detailed description of how to calibrate an assay, refer to EP7-P.13 Interference effects were assessed by Dose Response and
Section 6 of the instrument-specific operations manual. Paired Difference methods, at the medical decision level of the analyte.
QUALITY CONTROL Interfering
Interferent Concentration N
Target Observed
The following is the recommendation of Abbott Laboratories for quality Substance (U/L) (% of Target)
control. As appropriate, refer to your laboratory standard operating 30 mg/dL (513 µmol/L) 4 313.2 99.1
procedure(s) and/or quality assurance plan for additional quality control Bilirubin
60 mg/dL (1,026 µmol/L) 4 313.2 96.5
requirements and potential corrective actions.
62 mg/dL (0.62 g/L) 4 283.4 120.9
• Two levels of controls (normal and abnormal) are to be run every Hemoglobin
24 hours. 125 mg/dL (1.25 g/L) 4 283.4 143.8
• If more frequent control monitoring is required, follow the established 1,000 mg/dL (10.0 g/L) 4 302.0 96.8
Intralipid
quality control procedures for your laboratory. 2,000 mg/dL (20.0 g/L) 4 302.0 91.6
• If quality control results do not meet the acceptance criteria
defined by your laboratory, patient values may be suspect. Follow Bilirubin solutions at the above concentrations were prepared by addition
the established quality control procedures for your laboratory. of a bilirubin stock to human serum pools. Hemoglobin solutions at
Recalibration may be necessary. the above concentrations were prepared by addition of hemolysate to
human serum pools. Intralipid solutions at the above concentrations were
• Review quality control results and acceptance criteria following a prepared by addition of Intralipid to human serum pools.
change of reagent lot.

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SPECIFIC PERFORMANCE CHARACTERISTICS BIBLIOGRAPHY
(Continued) 1. Burtis CA, Ashwood ER, editors. Tietz Textbook of Clinical
Chemistry, 2nd ed. Philadelphia, PA: WB Saunders; 1994:813–8.
Precision
2. Amador E, Dorfman LE, Wacker WEC, et al. Serum lactic
The imprecision of the LD assay is ≤ 4.7% Total CV. Representative
dehydrogenase activity: an analytical assessment of current assays.
data from studies using CLSI protocol EP5-A14 are summarized below.
Clin Chem 1963;9:391.
Control Level 1 Level 2 3. US Department of Labor, Occupational Safety and Health
Administration. 29 CFR Part 1910.1030, Occupational Exposure to
N 80 80 Bloodborne Pathogens.
Mean (U/L) 162.6 422.1 4. US Department of Health and Human Services. Biosafety in
Microbiological and Biomedical Laboratories. HHS Publication
SD 1.94 1.54
Within Run (CDC), 4th ed. Washington, DC: US Government Printing Office,
%CV 1.2 0.4 May 1999.
SD 1.22 1.23 5. World Health Organization. Laboratory Biosafety Manual. Geneva:
Between Run World Health Organization, 2004.
%CV 0.8 0.3 6. Sewell DL, Bove KE, Callihan DR, et al. Protection of Laboratory
SD 0.00 3.84 Workers from Occupationally Acquired Infections; Approved
Between Day Guideline—Third Edition (M29-A3). Wayne, PA: Clinical and
%CV 0.0 0.9 Laboratory Standards Institute, 2005.
SD 2.30 4.32 7. Guder WG, Narayanan S, Wisser H, et al. List of analytes—
Total preanalytical variables. Annex In: Samples: From the Patient to the
%CV 1.4 1.0 Laboratory. Darmstadt, Germany: GIT Verlag; 1996:Annex 18–9.
Method Comparison 8. US Pharmacopeial Convention, Inc. General notices. In: US
Correlation studies were performed using CLSI protocol NCCLS Pharmacopeia National Formulary, 1995 ed. (USP 23/NF 18).
EP9-A.15 Rockville, MD: The US Pharmacopeial Convention, Inc; 1994:11.
Serum results from the LD assay on an AEROSET System were 9. Data on file at Abbott Laboratories.
compared with those from a commercially available modified Wacker 10. Sasse EA, Aziz KJ, Harris EK, et al. How to Define and Determine
methodology. Reference Intervals in the Clinical Laboratory; Approved Guideline
Serum results from the LD assay on an ARCHITECT c System were (C28-A). Villanova, PA: The National Committee for Clinical
compared with the LD assay on an AEROSET System. Laboratory Standards, 1995.
11. Passey RB, Bee DE, Caffo A, et al. Evaluation of the Linearity
AEROSET vs. ARCHITECT vs. of Quantitative Analytical Methods; Proposed Guideline (EP6-P).
Comparative Method AEROSET Villanova, PA: The National Committee for Clinical Laboratory
Standards, 1986.
N 75 97
12. Young DS. Effects of Drugs on Clinical Laboratory Tests, 4th ed.
Y - Intercept -5.466 -5.159 Washington, DC: AACC Press; 1995:3-372–3-377.
Correlation Coefficient 0.998 1.000 13. Powers DM, Boyd JC, Glick MR, et al. Interference Testing in
Slope 0.960 1.042 Clinical Chemistry; Proposed Guideline (EP7-P). Villanova, PA: The
National Committee for Clinical Laboratory Standards, 1986.
Range (U/L)* 25.3 to 912.5 71.0 to 4,451.3
14. Kennedy JW, Carey RN, Coolen RB, et al. Evaluation of Precision
*AEROSET Range Performance of Clinical Chemistry Devices; Approved Guideline
(EP5-A). Wayne, PA: The National Committee for Clinical
Laboratory Standards, 1999.
15. Kennedy JW, Carey RN, Coolen RB, et al. Method Comparison and
Bias Estimation Using Patient Samples; Approved Guideline (EP9-A).
Wayne, PA: The National Committee for Clinical Laboratory
Standards, 1995.

TRADEMARKS
AEROSET and ARCHITECT are registered trademarks of Abbott
Laboratories.
c System is a trademark of Abbott Laboratories.
All other trademarks, brands, product names, and trade names are the
property of their respective companies.

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ARCHITECT c SYSTEMS ASSAY PARAMETERS

Lactate Dehydrogenase Serum/Plasma—Conventional and SI Units

Configure assay parameters — General Configure assay parameters — SmartWash


● General о Calibration о SmartWash о Results о Interpretation о General о Calibration ● SmartWash о Results о Interpretation
Assay: LD Type: Photometric Version: † Assay: LD
Number: 1025
COMPONENT REAGENT / ASSAY WASH Volume Replicates
● Reaction definition о Reagent / Sample о Validity checks R1 TP000 0.5% AcidWash 345 1
Reaction mode: Rate up Cuvette Trig 10% Detergent B*** 345
Primary Secondary Read times
Wavelength: 340 / 380 Main: 21 – 28 *** Select “Detergent B” for software prior to Version 2.2.
Last required read: 28 Flex: 19 – 22
Absorbance range: ††† Color correction: ___ – ___
Sample blank type: None Configure assay parameters — Results
о General о Calibration о SmartWash ● Results о Interpretation
о Reaction definition ● Reagent / Sample о Validity checks Assay: LD Result units: U/L
R1 R2 Assay defaults:
Reagent: LD000 Reagent volume: 160 40 Low-Linearity: 8‡‡
Diluent: Saline Water volume: ___ ___ High-Linearity: 1984
Diluent dispense mode: Type 0 Dispense mode: Type 0 Type 0 Gender and age specific ranges:
Diluted Default GENDER AGE (UNITS) NORMAL EXTREME
Dilution name Sample sample Diluent Water Dilution factor dilution Either 0 – 130 (Y) 125 – 243
STANDARD : 3.2 ___ ___ ___ = 1:1.00 ●
1:5 : 20.0 3.2 80 ___ = 1:5.00 о
_________ : ___ ___ ___ ___ = о Configure result units
Assay: LD
Version: †
о Reaction definition о Reagent / Sample ● Validity checks Result units: U/L
Reaction check: None
Decimal places: 0 [Range 0 – 4]
Correlation factor: 1.0000
Intercept: 0.0000
Rate linearity %: 10

Configure assay parameters — Calibration


о General ● Calibration о SmartWash о Results о Interpretation
Assay: LD Calibration method: Factor
Factor: ‡
● Calibrators о Volumes о Intervals о Validity checks
Calibrator set: Calibrator level: Concentration:
None Blank: Water 0

Replicates: 3 [Range 1 – 3]

о Calibrators ● Volumes о Intervals о Validity checks


Calibrator: Diluted
Calibrator level Sample sample Diluent Water
Blank: Water 3.2 ___ ___ ___

о Calibrators о Volumes ● Intervals о Validity checks


Calibration intervals:
Full interval: 720 (hours)

о Calibrators о Volumes о Intervals ● Validity checks


Blank absorbance range: _____ – _____

† Due to differences in instrument systems and unit configurations, version numbers may vary.
††† c 8000 Absorbance range = 0.0000-1.0000; c 16000 Absorbance range = 0.0000-1.1700.
‡ The calibration factor for c 8000 is 13341; the calibration factor for c 16000 is 13920.
‡‡ The linear low value (Low-Linearity) is LOQ rounded up to the number of decimal places defined in the decimal places parameter field.

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AEROSET SYSTEM ASSAY PARAMETERS

Lactate Dehydrogenase Serum/Plasma—Conventional and SI Units

Assay Configuration: Outline Page


Assay Name Assay # Line
LD 25 A-Line
Quantitative Ranges
Min Text Min Panic-L L-Reference-H Panic-H Max Max Text
* 0.0* 0.0 125 243 0.0 0.0* *
8** L-Linear Range-H 1984
Reference Ranges*
Age Male Female
0 Year 0.0 – 0.0 0.0 – 0.0
0.0 – 0.0 0.0 – 0.0
0 Year
0.0 – 0.0 0.0 – 0.0
0 Year 0.0 – 0.0 0.0 – 0.0
Qualitative Ranges N/A

Assay Configuration: Base Page


Reaction Mode Wavelength-Prim/Sec Read time-Main/Flex Linearity %
RATE UP 340 / 380 21 – 28 / 19 – 22 10
Sample Blank Test Blank Read Time Abs Window Abs Limits
_____ ( ___ ) 0–0 0–0 0.0 – 1.0
S.Vol DS.Vol D.Vol W.Vol
Standard 3.2 0.0 0 0 Rgt Name/Pos
Dil 1 20.0 3.2 80 0 Diluent: DILUENT C–10*
Dil 2 3.2 0.0 0 0 Type# 0
Rgt Name/Pos R.Vol W.Vol Type#
Reagent 1 LD00061 – ___* 160 0 0
Reagent 2 LD00052 – ___* 40 0 0
Reaction Check Read Time – A/B Range Minimum
___________ 1–1/1–1 0.0 – 0.0 0.0
Factor/Intercept Decimal Places Units
1.0 / 0.0 0 U/L

Assay Configuration: Calibration Page


Calib Mode Factor Interval (H)
Factor 13341 720
Blank/Calib Replicates Span Span Abs Range
3/0 BLK – 1 0.0 – 0.0
Sample S.Vol DS.Vol D.Vol W.Vol Blk Abs Range
BLK Water 3.2 0.0 0 0 0.0 – 0.0
C1 2.0 0.0 0 0 Cal Deviation
C2 2.0 0.0 0 0 0.0

Assay Configuration: SmartWash Page


Rgt Probe
Reagent Wash Vol
TP00061 AcidW 345
DIG0051 Water 345
DIG0012 Water 345
Cuvette
Assay Name Wash Vol
— — —
Sample Probe
Wash

Refer to Assay Configuration in Section 2 of the AEROSET System Operations Manual for information regarding assay parameters.
* User defined or instrument defined.
** The linear low value (L-Linear Range) is LOQ rounded up to the number of decimal places defined in the decimal places parameter field.

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