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High-density lipoprotein (HDL)
Web： http://htidiagnostics.com HT-H233E-40 HT-H233E-340 Reagent Set
HT-H233E-80 HT-H233E-540
High Technology, Inc. HT-H233E-320

INTENDED USE LIMITATIONS

For the quantitative determination of high-density lipoprotein (HDL) in human serum or plasma on automated analyzer. 1. Anticoagulants containing citrate should not be used.
For in vitro diagnostic use only. 2. Protect the reagents from direct sunlight.
3. Store the reagents as per instructions.
SUMMARY AND EXPLANATION OF THE TEST 4. Samples with values greater than 150 mg/dl must be diluted 1:1 with saline and re-assayed. Multiply the result by
Cholesterol is a fatty substance found in blood, bile and brain tissue. It serves as a precursor to bile acids, steroids and two.
vitamin D. The concentration of total cholesterol in serum has been associated with metabolic, infectious and coronary
heart diseases. In the plasma, cholesterol is transported by three lipoproteins: high-density lipoprotein (HDL- CALIBRATION
Cholesterol), low density lipoprotein (LDL-Cholesterol), and very low density lipoprotein (VLDL-Cholesterol). The Direct HDL/LDL Cholesterol Calibrator is required for calibration. Calibrate with each bottle change or lot change
or if control results are found to be out of range. The value of the Direct HDL/LDL calibrator was assigned by
The role of HDL particles in lipid metabolism is primarily the uptake and transport of cholesterol from peripheral tissue procedures traceable to the National Reference System for Cholesterol (NRS/CHOL). Refer to Direct HDL/LDL
to the liver. This process is known as reverse cholesterol transport and has been proposed as a cardio protective Cholesterol calibrator kit package insert for instructions. If control results are found to be out of range, the procedure
mechanism. Low HDL-C levels have repeatedly been associated with an increased risk of coronary heart disease and should be recalibrated.
coronary artery disease. Thus the determination of serum HDL cholesterol has been recognized as a useful tool in
identifying high-risk patients. QUALITY CONTROL
Reliability of test results should be routinely monitored with control materials that reasonably emulate performance of
The CDC reference method for HDL cholesterol uses ultracentrifugation followed by chemical precipitation to separate patient specimens. 10 Quality control materials are intended for use only as monitors of accuracy and precision. Any
HDL from other lipoproteins, followed by cholesterol measurement using a modified Abell-Kendall assay. This method HDL/LDL controls would be suitable for use with this assay. The National Cholesterol Education Program (NCEP) Lipid
is considered too time consuming and labor intensive for use in routine analysis. Historically, most laboratories have Standardization Panel (LSP) recommends two levels of controls, one in the normal range (35-65 mg/dl) and one near
used one of several methods for the selective precipitation and removal of LDL and VLDL, followed by the enzymatic the concentrations for decision making (<35 mg/dl). An acceptable range of HDL cholesterol values should be
measurement of HDL-C in the supernatant fraction. Since almost all of these methods required manual separation established for the controls by repeat analysis. The recovery of control values within the appropriate range should be
steps, HDL cholesterol determinations could not be fully automated. Also, the dilution of the sample resulted in an the criteria used in evaluation of future assay performance. Quality control materials are intended for use only as
enzymatic determination of cholesterol with low sensitivity. As a result, the routine determination of HDL cholesterol monitors of accuracy and precision. Controls should be run with every working shift in which HDL -C assays are
has suffered form both long turnaround times and poor reproducibility. performed. It is recommended that each laboratory establish their frequency of control determination. Quality control
requirements should be performed in conformance with local, state, and/or Federal regulations or accreditation
PRINCIPLE requirements.
The Direct HDL Cholesterol assay is a homogeneous method for directly measuring serum HDL-C levels without the
need for any off-line pretreatment or centrifugation steps. The method is in a two-reagent format. The first reagent EXPECTED VALUES
stabilizes LDL, VLDL, and chylomicrons. The second reagent contains PEG modified enzymes that selectively react The expected values for serum HDL cholesterol are as follows:
with the cholesterol present in the HDL particles. Consequently, only the HDL cholesterol is subject to cholesterol < 40 mg/dl Low
measurement. > 60 mg/dl High
It is recommended that each laboratory establish its own range of expected values, since differences exist between
REAGENT COMPOSITION instruments, laboratories, and local populations.
1. Direct HDL Cholesterol Reagent 1: According to the NCEP, HDL values greater than or equal to 35 mg/dl are considered desirable, and values greater
Magnesium chloride 100mM, Aminoantipyrene 1 mmol/L, buffer, pH 7.0+0.1, preservative. than or equal to 60 mg/dl are considered to offer some protection against coronary heart disease. Values below 35
2. Direct HDL Cholesterol Reagent 2: mg/dl are considered to be a significant independent risk factor for coronary heart disease.
Peroxidase from Horseradish (POD) 4KU/L, Cholesterol Oxidase from Nocardia sp. (PEG-CO) 1 KU/L, Cholesterol
Esterase from Pseudomonas (PEG-CE) 1KU/L, N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline (HDAOS) 0.3g/L, PERFORMANCE CHARACTERISTICS
buffer, pH 7.0+0.1, surfactant, preservative. Assay Range: 2 – 150 mg/dl
Accuracy: Accuracy of the Direct HDL Cholesterol Reagent method was verified by comparison to the Designated
WARNINGS AND PRECAUTIONS Comparison Method (ultracentrifugation, chemical precipitation and enzymatic cholesterol analysis which has been
1. For in vitro diagnostic use only. standardized to the Abell-Kendall method)10 and another manufacturer’s lyophilized Direct HDL Cholesterol method to
2. Exercise the normal precautions required for the handling of all laboratory reagents. Pipetting by mouth is not the Designated Comparison Method produced the following results:
recommended for any laboratory reagent.
3. All specimens used in this test should be considered potentially infectious. Universal precautions as they apply to Studies comparing the Direct HDL Cholesterol method to Pointe Scientific AutoHDL Cholesterol Reagent Set on
your facility should be used for handling and disposal of materials during and after testing. Hitachi 717 Analyzer produced the following results:
4. Do not use the reagent after the expiration date printed on the kit. Method Direct HDL Cholesterol AutoHDL Cholesterol
N 52 52
STORAGE AND STABILITY Mean HDL Cholesterol 46 48.3
Store the reagent set at 2 – 8 °C. The reagent is stable until the expiration date indicated on the bottle label. Range (mg/dl) 22.3-96.8 21.3-92.1
Regression Analysis y=0.93x+0.73mg/dl
SPECIMEN COLLECTION AND STORAGE Correlation Coefficient r=0.916
Serum, EDTA-treated or heparinized plasma are the recommended specimens.
Serum: Collect whole blood by venipuncture and allow to clot. Centrifuge and remove the serum as soon as possible Precision: Within Day precision for the Direct HDL Cholesterol Reagent was determined on Hitachi 717 following a
after collection (within 3 hours). modification of NCCLS document EP5-T2. Within Day precision studies produced the following results:
Plasma: Specimens may be collected in EDTA or heparin. Centrifuge and remove the plasma as soon as possible
after collection (within 3 hours). If not analyzed promptly, specimens may be stored at 2-8°C for up to 1 week. If Within Day
specimens need to be stored for more than 1 week, they may be preserved at less than -20°C for up to 1 month. For Sample 1 Sample 2
storage periods of 1 month to 2 years, samples should be preserved at -70°C. N 25 25
Mean HDL Cholesterol 38.1 83.5
INTERFERENCES Standard Deviation (mg/dl) 1.54 1.94
All interference studies were conducted according to the procedures recommended in NCCLS guideline NO. EP7 -P for Correlation of Variation (%) 4.0 2.3
interference testing in clinical chemistry. Hemoglobin levels up to 100 mg/dl, Triglyceride levels up to 1800 mg/dl and Day to Day
Bilirubin levels up to 20 mg/dl were found to exhibit negligible interference (<5%) on this method. Samples with levels Day to Day precision for the Direct HDL Cholesterol Reagent was also determined on Hitachi 717 following a
of interfering substances higher than the upper limits should be diluted with physiological saline before assaying. Refer modification of NCCLS document EP5-T2. Day to Day precision studies produced the following results:
to the work of Young for a review of drug effects on serum HDL cholesterol levels.
Sample 1 Sample 2
MATERIALS PROVIDED N 25 25
1. Direct HDL Cholesterol Reagent 1 (Ready to use) Mean HDL Cholesterol 38.2 83.4
2. Direct HDL Cholesterol Reagent 2 (Ready to use) Standard Deviation (mg/dl) 1.74 2.2
Correlation of Variation (%) 4.5 2.6
MATERIALS REQUIRED BUT NOT PROVIDED
1. Direct HDL/LDL Cholesterol Calibrator Sensitivity: The analytical sensitivity of the Direct HDL Cholesterol Reagent was determined on Hitachi 717 as 2 mg/dl
2. Lipid Control Set of HDL Cholesterol.
3. Automated clinical chemistry analyzer capable of measuring absorbance at 700 and 600 nm.
Manufactured by
PROCEDURE High Technology, Inc.
Below is a general example of the Direct HDL Cholesterol test procedure for an automated analyzer. All analyzer 109 Production Rd, Walpole, MA 02081, USA
applications should be validated in accordance with NCEP and CLIA recommendations. Tel: 1-508-660-2221
1. Use 4 ul sample with 300 ul of Direct HDL Cholesterol Reagent 1. E-mail: [email protected]
2. Equilibrate to 37°C for 5 minutes. www.htmed.com, http://htidiagnostics.com
3. Add 100 ul of Direct HDL Cholesterol Reagent 2. Manufactured by Used by
4. Equilibrate to 37°C for 5 minutes.
5. Measurement (Absorb. Difference between 700nm & 600 nm)
Catalog number In vitro diagnostic
CALCULATIONS
(A = Absorbance)
Temperature Authorized representative in
Δ A (patient) × Concentration of calibrator = Concentration of
limitation the European Community
Δ A (standard) (mg/dl) patient (mg/dl)

Example: Lot number Consult instructions for use

Δ A (patient) = 0.40, Δ A (calibrator) = 0.32, Concentration of calibrator = 53 mg/dl.
This product conforms to the directive 98/79/EC (IVD-directive)
0.40 × 53 = 66.3 mg/dl
0.32
To convert from conventional units to SI units, multiply the conventional units by 0.02586.
mg/dl × 0.02586 = mmol/L HDL cholesterol

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