Download name and intended use

within a week or so of the estimated date of
conception.
SPECIMEN COLLECTION AND HANDLING
to get the most reliable and
consistent results.
3.
VISUAL COLOR TEST FOR
EARLY PREGNANCY
Catalog No. MYM/US-V_HCG
For In Vitro Diagnostic Use Only
NAME AND INTENDED USE
The MYM hCG HP-501 is designed for in vitro
qualitative detection of Human Chorionic Gonadotropin
(hCG) in patient’s urine and serum. It is a visual early
pregnancy test. (For in vitro diagnostic use)
PRINCIPLE OF THE ASSAY
Urine specimen
UBI HCG- MYM is a solid phase enzyme-linked
immunosorbent assay (ELISA) based on the
Collect urine in a container without any
preservative. The preferred specimen is the
first morning urine because it usually has the
maximum concentration of hCG. Specimen
that cannot be assayed immediately must be
stored at 2-8oC for 24 hrs. or frozen up to six
months. Avoid repeated freezing and thawing
of urine sample.
sandwich principle. The wells are coated with
a monoclonal antibody directed toward a
unique antigenic site on the hCG molecule.
The samples are incubated in the wells with
enzyme conjugate, which is a second
monoclonal antibody, directed toward a
different regions of hCG molecule and
conjugated with horseradish peroxidase, to
form a sandwich complex bound to the well.
Unbound conjugate are then washed off with
tap water. The amount of bound peroxidase is
proportional to the concentration of the hCG
present in the sample. Upon addition of the
substrate and chromogen, the intensity of blue
color developed is proportional to the
concentration of hCG in the sample.
Serum Specimen
Collect blood by venipuncture. Allow it to clot
and separate the serum by centrifugation at
room temperature. If sera cannot be assayed
immediately, they can be stored at 2-8C for a
week or frozen for up to six months. Avoid
repeated freezing and thawing of serum
sample.
Chorionic Gonadotrpopin (hCG) is a
gycoprotein hormone normally produced by the
placenta during pregnancy. The hormone is
present in blood and urine around seven to
thirteen days following implantation of the
fertilized ovum. hCG increases in concentration
at a geometric rate, reaching a peak near the
end of the first trimester. High concentrations
are observed throughout pregnancy. After
parturition, concentration declines rapidly, and
becomes undetectable within a few days.
Structurally intact hCG molecules consist of
two non-covalently linked poly-peptide
subunits, the alpha and beta chain subunits.
The alpha subunit is similar to that of other
glycoprotein hormones, such as Thyroid
Stimulating Hormone (TSH), Luteinizing
Hormone(LH), and Follicle Stimulating
Hormone(FSH). The differences in the amino
acid sequences of the beta subunits of the
respective hormones accounts for their target
organ specificities and immunochemical
distinctiveness. Monoclonal antibodies
produced from hybridomas represent an
essentially unlimited supply of homogeneous
antibody of precisely defined specificity.
Monoclonal antibodies which react with unique
sites on the beta chain of the hCG molecule are
essential for differentiation between hCG and
LH, FSH and TSH. Assay for the beta subunit
has permitted the detection of pregnancy
ASSAY PROCEDURE
1.
2.
3.
4.
5.
6.
7.
8.
9.
SUMMARY AND EXPLANATION OF TEST
WARNING AND PRECAUTION



MYM kit is designed for in vitro use
only.
The Reference and Negative Controls
are formulated in a salt-protein
matrix. These Controls can be
applied to both urine or serum
specimens.
The components in this kit are
intended for use as an integral unit .
The components of different lots
should not be mixed.
MATERIALS PROVIDED
1.
2.
3.
4.
5.
STORAGE AND STABILITY

Store the kit at 2-8C in a
refrigerator.
Keep MAGIWEL in a dry bag with
7.
8.
desiccants.

The unopened reagents are
stable until expiration of the kit.
Solution A and Solution B should
be colorless; if solutions turn
blue, it must be replaced. Do
not expose test reagents to
strong light during storage or
usage.
Secure the desired number of coated
wells in the holder. Mark data sheet
with sample identification.
Dispense 50 uL of urine or serum or
controls into appropriate wells.
Dispense 50 uL of enzyme conjugate
into each wells
Incubate for 5 minutes or longer at
room temperature.
Remove incubation mixture by
flicking the plate into sink.
Rinse the wells 5 times with running
tap water.
Dispense 50 ul of Solution A and 50
ul of Solution B into each well.
Incubate 5 minutes or longr at room
temperature.
Observe blue color development and
mark data.
(UBI HCG MAGIWEL Cat No. HP-501)
6.

Use a new disposable pipet tip for
each specimen.
Microwell Strips: Anti-hCG
monoclonal antibodies coated wells.
Enzyme Conjugate (6 mL): Anti-hCG
monoclonal antibody conjugated to
horseradish peroxidase.
Solution A : Buffer solution containing
peroxide.
Solution B: Tetramethybenzidine.
Negative Control (0.75 mL): Saltprotein matrix with 0.01% proclin.
Cut off control(0.75 mL): Calibrated
to 25 mIU hCG/mL with preservative
(0.01% proclin).
Positive control: Calibrated to 100
mIU hCG/mL with preservative
(0.01% proclin).
Well Holder: For securing individual
wells.
PREPARATION FOR ASSAY
1.
2.
Before beginning the test, bring all
samples and reagents to room
temperature (20-25C) and gently
mix.
Have all reagents and samples ready
before the start of the assay. Once
the test has begun it must be
performed without any interruptions
RESULTS AND INTERPRETATION



Negative Results
No color indicates a nondetectable
amount of hCG in the sample.
Positive Results
Blue color the same as or more
intense than the Cut off Control (25
mIU/mL) indicates presence of hCG
in the sample.
Indeterminate Result
Less blue color than the 25 mIU
hCG/mL Cut off Control indicates hCG
concentration less than 25 mIU/mL.
Another sample taken from the
patient at least 48 hours later should
be reassayed side by side in the
repeat assay. The Cut off, Positive
and Negative Controls run with
patient sample establish the validity
of a test. The control concentrations
of hCG were arbitrarily selected. To
be positive or negative for pregnancy.
Conditions such as hydatidiform
mole, choriocarcinoma, trophoblastic
or nontrophoblastic neoplasms may
be associated with the presence of
hCG in the sample.
EXPECTED VALUES AND LIMITATIONS



hCG is normally not detected in the
urine or serum of healthy men and
nonpregnant women by UBI HCG
MAGIWEL. The urine hCG
concentrations of pregnant women
begin to rise as soon as the first week
following implantation, reach 40
mIU/mL ten days after concenption,
and reach a maximum concentration
(in excess of 100,000 mIU/mL)
during the latter part of the first
trimester of pregnancy.
During the first six weeks of
pregnancy, serum hCG
concentrations have a doubling time
of approximately 2 days. Following
delivery hCG concentrations rapidly
decrease and usually return to
normal in several days.
High levels of hCG may persist after
hCG injections in patients undergoing
infertility therapy. HCG may be
present in very low levels in ectopic
pregnancies while conditions like
choriocarcinoma, trophoblastic or
nontrophoblastic neoplasms, or
hydatidiform mole may result in high
concentrations of hCG.
PERFORMANCE CHARACTERISTICS
Sensitivity
The analytical sensitivity of the MAGIWEL test
has been set at 25 mIU/mL based on the 1st
international Standard (WHO 1st IRP)
Interference
The addition of 200 μg/ml of each of the
following compounds does not interfere with
MAGIWEL test for hCG: acetaminophen,
ascorbic acid, acetylsalicylic acid, atropine,
gentisic acid, methadone, and phenothiazine.
Abnormally elevated levels of protein, ketones,
bilirubin or hemoglobin do not affect the essay.
Accuracy
Analysis of Urine Samples: Urine samples
from known non-pregnant subjects (N=30) are
spiked with known amountsof hCG to a final
concentration of 12.5, 25 and 50 mIU/mL.
HCG Added
(mIU/mL)
Number of
Assay
Positive
Negative
Indeterminate
Sensitivity(%)
0
12.5
25
50
30
0
30
0
30
3
2
25
30
28
0
2
93
30
30
0
0
100
Analysis of Serum Samples: Serum
samples from known non-pregnant subject
(N=20) are spiked with known amounts of hCG
to a concentration of 12.5, 25, 50 mIU/mL.
Specificity
250 mIU/mL
hFSH 1,000 mIU/mL
hTSH 1,000 IU/mL
Negative
Negative
Negative
HCG Added
(mIU/mL)
Number of
Assay
Positive
Negative
Indeterminate
Sensitivity(%)
0
20
0
20
0
12.5
25
50
20
0
20
0
20
19
0
1
95
20
20
0
0
100
Qualitative
EIA
Quantitative
EIA
100
100
100
44
44
44
56
56
56
Each laboratory should utilize internal controls
at several levels to monitor assay
performance. The control should be treated as
unknown. Results obtained should be in
agreement with the assigned value of the
control. The controls can be obtained from
commercially available sources, but should not
contain sodium azide as preservative.
REFERENCE
1.
3.
4.
Correlation with EIA Methods
Serum samples from 100 females are analyzed
by MAGIWEL visual test and with other
accepted EIA methods as following:
UBI
MAGIWEL
Quality Control
2.
The immunological specificity of MAGIWEL test
was determined from cross reaction with
known quantities of purified preparation of
Luteinizing Hormone (hLH), Follicle Stimulating
Hormone (FSH) and Thyroid Stimulating
Hormone (hTSH).
hLH
Methods
Number
of Assay
No.of
Samples
with hCG
> 25
mIU/mL
No.of
Samples
with hCG
< 25
mIU/mL
5.
Lenton, E. L. Neal and R. Sulaimlan.
Plasma concentrations of human
gonadotropin from time of
implantation until second week of
pregnancy. Fertility and Sterility 37:
773, 1982.
Braunstein, G.D. First-trimester
chorionic gonadotropin
measurements as an aid in the
diagnosis of early pregnancy
disorders Am. J. Obst. Gynec.
131:25, 1978.
Kosasa, T.S. Measurement of human
chorionic gonadotropin. J. Reprod.
Med. 26:201, 1981.
Catt, K.J., Dufau, M.L., and
Vaitukaitis, J.L., Appearance of hCG
in Pregnancy plasma following the
initiation of implantation of the
blastrocyst. J.Clin.Endocr. Metab.
40:537, 1975.
Uotila, M., Ruoslahti, E., and Engvall,
E., Two-site sandwich enzyme
immunoassay with monoclonal
antibodies to human alpha
fetoprotein. J. Immunol Methods 42:
11, 1981.
6. Braunstein, G.D., Rasor,J.,
Adler,D.,Danzer,H.,and
Wade,M.E.Serum human chorionic
gonadotropin levels throughout
normal pregnancy.
Am.J.Obstet.Gynecol. 126:678, 1976.
7. Batzer,F. Hormonal evaluation of
early pregnancy. Fertility and
Sterility 34:1, 1980.
8. Dipietro, D.L., Ectopic pregnancy:
interpreting HCG levels. Laboratory
Management 19:11, 1981
9. Engvall, E., and Perlmann, P.,
Enzyme linked immunosorbent assay
(ELISA) quantitative assay of
immunoglobulin. Immunochemistry
8:871, 1971.
10. Bangham, D.R. and Storring, P.L.
Standardization of human choironic
gonadotropin, hCG subunits, and
pregnancy test. Lancet 1:390, 1982.
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