MAGLUMI free β-HCG (CLIA)
Ref: 130214002M
INTENDED USE
The kit has been designed for the quantitative determination of
free β-chains of human chorionic gonadotropin (free β-HCG) in
human serum.
The method can be used for samples over the range of 1.1-200
ng/ml.
The test has to be performed on the Fully-auto
chemiluminescence immunoassay (CLIA) analyzer MAGLUMI
(Including Maglumi 600,Maglumi 1000,Maglumi 1000 Plus,
Maglumi 2000,Maglumi 2000 Plus,Maglumi 3000 and Maglumi
4000).
SUMMARY AND EXPLANATION OF THE TEST
Human chorionic gonadotropin (HCG) is a dimeic glycoprotein with
a molecular weight of 37,000 Dalton consisting of an α and a
β-subunit. The α–subunit of HCG is nearly identical with the
analogous subunit of the pituitary hormones LH, FSH, and TSH.
HCG is produced by the syncytiotrophoblast of the placenta and
can be detected in maternal serum immediately upon nidation.
HCG is responsible for maintain the function of the ovarian corpus
luteum. In normal pregnancy, HCG concentrations rapidly increase
with a doubling rate of approximately two days. Pathologically
decreased values indicate abnormal pregnancy (e.g. ectopic
pregnancy). A sudden fall of HCG levels before the 8th -10th week
of gestation should be considered as a serious indicator of
imminent abortion.
If pregnancy can be excluded, HCG levels above normal are
highly likely to indicate the presence of malignant neoplasms,
particularly germ-cell tumors (e.g. testicular tumors, chorionic or
ovarian carcinoma. For use as a tumor marker, the test system
has to recognize both HCG and β-HCG, since testicular and
chorionic carcinomas mainly secrete the free β-chains rather than
the intact HCG molecule
PRINCIPLE OF THE TEST
Sandwich immunoluminometric assay:
Use an anti-free-β-HCG monoclonal antibody to label ABEI, and
use another monoclonal antibody to label FITC. Sample,
Calibrators, or Control with ABEI Label, FITC Label and magnetic
microbeads coated with anti-FITC are mixed thoroughly and
incubated at 37℃, forming a sandwich; After sediment in a
magnetic field, decant the supernatant, cycle washing for 1 time.
Subsequently, the starter reagents are added and a flash
chemiluminescent reaction is initiated. The light signal is measured
by a photomultiplier as RLU within 3 seconds and is proportional to
the concentration of free-β-HCG present in samples.
KIT COMPONENTS
Material Supplies: Reagent Integral for 100 determinations
Nano magnetic microbeads: TRIS buffer,
1.2% (W/V), 0.2%NaN3, coated with sheep
anti- FITC polyclonal antibody: 2.5ml
Calibrator Low: bovine serum, 0.2%NaN3: 2.5ml
Calibrator High: bovine serum, 0.2%NaN3: 2.5ml
FITC label: anti-free-β-HCG monoclonal
antibody labeled FITC contains BSA,
0.2%NaN3: 10.5ml
ABEI label: anti-free-β-HCG monoclonal
antibody labeled ABEI, contains BSA,
0.2%NaN3: 10.5ml
All reagents are provided ready-to-use.
Reagent Vials in kit box
Internal Quality Control: containing BSA,
0.2%NaN3. (target value refer to Quality
Control Information date sheet): 2.0ml
Internal quality control is only applicable with MAGLUMI system.
Instructions for use and target value refer to Quality Control
Information date sheet. User needs to judge results with their own
standards and knowledge.
Accessories Required But Not Provided
MAGLUMI Reaction Module REF: 630003
MAGLUMI Prenatal Screening Software REF:1301221
MAGLUMI Starter 1+2 REF: 130299004M
MAGLUMI Wash Concentrate REF: 130299005M
MAGLUMI Light Check REF: 130299006M
Preparation of the Reagent Integral
Before the sealing is removed, gentle and careful horizontal
shaking of the Reagent Integral is essential (avoid foam formation!)
Remove the sealing and turn the small wheel of the magnetic
microbeads compartment to and fro, until the colour of the
suspension has changed into brown. Place the Integral into the
reagent area and let it stand there for 30 min. During this time, the
magnetic microbeads are automatically agitated and completely
resuspended.
Do not interchange integral component from different
reagents or lots!
Storage and Stability
Sealed: Stored at 2-8℃ until the expiry date.
Opened: Stable for 4 weeks. To ensure the best kit performance,
it is recommended to place opened kits in the refrigerator if it’s not
going to be used on board during the next 12 hours.
CALIBRATION AND TRACEABILITY
1)Traceability
To perform an accurate calibration, we have provided the test
calibrators standardized against the WHO Reference Reagent
Human Chorionic Gonadotrophin, Beta Subunit (Purified) 99/650
2) 2-Point Recalibration
Via the measurement of calibrators, the predefined master curve is
adjusted (recalibrated) to a new, instrument-specific measurement
level with each calibration.
3) Frequency of Recalibration
After each exchange of lot (Reagent Integral or Starter
Reagents).
Every 2 weeks and/or each time a new Integral is used
(recommendation).
After each servicing of the Fully-auto chemiluminescence
immunoassay (CLIA) analyzer MAGLUMI.
If controls are beyond the expected range.
The room temperature has changed more than 5 ℃
(recommendation).
SPECIMEN COLLECTION AND PREPARATION
Sample material: serum
Collect 5.0ml venous blood into Blood Collection Tube. Standing at
room temperature,centrifuging, separating serum part.
The serum sample is stable for up to 12 hours at 2-8℃. More than
12 hours, please packed, -20 ℃ can be stored for 30 days.
Avoid repeated freezing and thawing, the serum sample can be
only frozen and thawed two times. Stored samples should be
thoroughly mixed prior to use (Vortex mixer).
Please ask local representative of SNIBE for more details if you
have any doubt.
Vacuum Tubes
(a) Blank tubes are recommended type for collecting samples.
(b) Please ask SNIBE for advice if special additive must be used in
sample collecting.
Specimen Conditions
Do not use specimens with the following conditions:
(a) heat-inactivated specimens;
(b) Cadaver specimens or body fluids other than human
serum;
(c) Obvious microbial contamination.
Use caution when handling patient specimens to prevent
cross contamination. Use of disposable pipettes or pipette tips
is recommended.
Inspect all samples for bubbles. Remove bubbles with an
applicator stick prior to analysis. Use a new applicator stick for
each sample to prevent cross contamination.
Serum specimens should be free of fibrin, red blood cells or
other particulate matter.
Ensure that complete clot formation in serum specimens has
taken place prior to centrifugation. Some specimens,
especially those from patients receiving anticoagulant or
thrombolytic therapy, may exhibit increased clotting time. If the
specimen is centrifuged before a complete clot forms, the
presence of fibrin may cause erroneous results.
Preparation for Analysis
+ Patient specimens with a cloudy or turbid appearance must be
centrifuged prior to testing. Following centrifugation, avoid the
lipid layer (if present) when pipetting the specimen into a
sample cup or secondary tube.
+ Specimens must be mixed thoroughly after thawing by low
speed vortexing or by gently inverting, and centrifuged prior to
use to remove red blood cells or particulate matter to ensure
consistency in the results. Multiple freeze-thaw cycles of
specimens should be avoided.
+ All samples (patient specimens or controls) should be tested
within 3 hours of being placed on board the MAGLUMI
System. Refer to the SNIBE service for a more detailed
discussion of onboard sample storage constraints
Storage
If testing will be delayed for more than 8 hours, remove serum
from the serum separator, red blood cells or clot. Specimens
removed from the separator gel, cells or clot may be stored up
to 12 hours at 2-8°C.
Specimens can be stored up to 30 days frozen at -20°C or
colder.
Shipping
Before shipping specimens, it is recommended that specimens
be removed from the serum separator, red blood cells or clot.
When shipped, specimens must be packaged and labeled in
compliance with applicable state, federal and international
regulations covering the transport of clinical specimens and
infectious substances. Specimens must be shipped frozen (dry
ice). Do not exceed the storage time limitations identified in this section of the package insert.
WARNING AND PRECAUTIONS FOR USERS
+ For use in IN-VITRO diagnostic procedures only.
+ Package insert instructions must be carefully followed.
Reliability of assay results cannot be guaranteed if there are
any deviations from the instructions in this package insert
Safety Precautions
CAUTION: This product requires the handling of human
specimens.
The calibrators in this kit are prepared from bovine serum
products. However, because no test method can offer
complete assurance that HIV, Hepatitis B Virus or other
infectious agents are absent; these reagents should be
considered a potential biohazard and handled with the same
precautions as applied to any serum or plasma specimen.
All samples, biological reagents and materials used in the
assay must be considered potentially able to transmit
infectious agents. They should therefore be disposed of in
accordance with the prevailing regulations and guidelines of
the agencies holding jurisdiction over the laboratory, and the
regulations of each country. Disposable materials must be
incinerated; liquid waste must be decontaminated with
sodium hypochlorite at a final concentration of 5% for at
least half an hour. Any materials to be reused must be
autoclaved using an overkill approach. A minimum of one
hour at 121℃ is usually considered adequate, though the
users must check the effectiveness of their decontamination
cycle by initially validating it and routinely using biological
indicators.
It is recommended that all human sourced materials be
considered potentially infectious and handled in accordance
with the OSHA Standard on Bloodborne Pathogens13.
Biosafety Level 214 or other appropriate biosafety practices
should be used for materials that contain or are suspected
of containing infectious agents.
This product contains Sodium Azide; this material and its
container must be disposed of in a safe way.
Safety data sheets are available on request
Handling Precautions
Do not use reagent kits beyond the expiration date.
Do not mix reagents from different reagent kits.
Prior to loading the Reagent Kit on the system for the first time,
the microbeads requires mixing to re-suspend microbeads
that have settled during shipment.
For microbeads mixing instructions, refer to the KIT
COMPONENTS, Preparation of the Reagent Integral section
of this package insert.
To avoid contamination, wear clean gloves when operating
with a reagent kit and sample.
Over time, residual liquids may dry on the kit surface, please
pay attention the silicon film still exists on the surface of the kit.
For a detailed discussion of handling precautions during
system operation, refer to the SNIBE service information.
TEST PROCEDURE
To ensure proper test performance, strictly adhere to the operating
instructions of the Fully-auto chemiluminescence immunoassay
(CLIA) analyzer MAGLUMI. Each test parameter is identified via a
RFID tag on the Reagent Integral. For further information please
refer to the Fully-auto chemiluminescence immunoassay (CLIA)
analyzer MAGLUMI Operating Instructions.
Sample, calibrator: 40μl
FITC Label: +80μl
ABEI Label: +80μl
Nano magnetic microbeads: +20μl
Incubation: 15 min
Cycle washing: 400μl
Measurement: 3s
DILUTION
Sample dilution by analyzer is not available in this reagent kit
Samples with concentrations above the measuring range can be
diluted manually. After manual dilution, multiply the result by the
dilution factor.
Please choose applicable diluents or ask SNIBE for advice before
manual dilution must be processed.
QUALITY CONTROL
Observe quality control guidelines for medical laboratories
Use suitable controls for in-house quality control. Controls
should be run at least once every 24 hours when the test is in
use, once per reagent kit and after every calibration. The
control intervals should be adapted to each laboratory’s
individual requirements. Values obtained should fall within the
defined ranges. Each laboratory should establish guidelines
for corrective measures to be taken if values fall outside the
range.
LIMITATIONS OF THE PROCEDURE
1) Limitations
A skillful technique and strict adherence to the instructions are
necessary to obtain reliable results. Bacterial contamination of
samples or repeated freeze-thaw cycles may affect the test results.
Assay results should be utilized in conjunction with other clinical
and laboratory data to assist the clinician in making individual
patient management decisions.
2) Interfering Substances
No interference with test results is seen by concentrations of
bilirubin<0.125mg/ml, haemoglobin<16mg/dl or triglycerides<
12.5mg/ml.
3) HAMA
Patient samples containing human anti-mouse antibodies (HAMA)
may give falsely elevated or decreased values. Although
HAMA-neutralizing agents are added, extremely high HAMA
serum concentrations may occasionally influence results.
4) High-Dose Hook
No high-dose hook effect was seen for free-β-HCG concentrations
up to 4000ng/ml.
RESULTS
1) Calculation of Results
The analyzer automatically calculates the free β-HCG
concentration in each sample by means of a calibration curve
which is generated by a 2-point calibration master curve procedure.
The results are expressed in ng/ml. For further information please
refer to the Fully-auto chemiluminescence immunoassay (CLIA)
analyzer MAGLUMI Operating Instructions.
2) Interpretation of Results
The result of free β-HCG (Prenatal Screening) assay is
calculated using the specific MAGLUMI Prenatal Screening
software (REF:1301221). Please check the software for
reference result.
Results may differ between laboratories due to variations in
population and test method. If necessary, each laboratory
should establish its own reference range.
PERFORMANCE CHARACTERISTICS
1) Precision
Intra-assay coefficient of variation was evaluated on 3 different
levels of control serum repeatedly measured 20 times in the same
run, calculating the coefficient of variation.