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Idiomas disponibles
Idiomas disponibles
Printed For Final Review: 10/10/03
ECO Number: 27341
7.
COLLECTION AND PREPARTION OF SPECIMENS
One hundred microliters, in duplicate, of either EDTA plasma or serum are required for
the assay. A fasting specimen is recommended.
For serum: collect blood by venipuncture in a 5 or 10 mL evacuated glass tube. Allow
the blood to clot at room temperature (20-25°C). Centrifuge for 15 minutes at 760 x g*
to obtain hemolysis-free sera. No additives or preservatives are required to maintain
integrity of the sample. All plastics, glassware or other material coming into contact with
the specimen should be entirely free of any contamination.
For plasma: collect blood by venipuncture in a 5 or 10 mL evacuated glass tube. EDTA
(7.2 mg/5 mL blood) is used as anticoagulant. Centrifuge for 15 minutes at 760 x g.* No
additives or preservatives are required to maintain integrity of the sample. All plastics,
glassware or other material coming into contact with the specimen should be entirely
free of any contamination.
Serum or plasma samples should be stored frozen.
repeatedly frozen and thawed.
8.
EQUIPMENT AND MATERIALS REQUIRED, BUT NOT SUPPLIED
8.1 Disposable borosilicate glass tubes, 12 x 75 mm.
8.2 Temperature controlled centrifuge to accommodate 12 x 75 mm tubes (20-
25°C).
8.3 Gamma scintillation counter capable of counting iodine-125.
8.4 Vortex.
8.5 Pipetting devices:
a.
Micropipettors calibrated to deliver 100 µL, 200 µL, and 500 µL.
b.
Repeating dispensers, calibrated to deliver 200 µL and 500 µL.
8.6 Refrigerator at 2-8°C or crushed ice bath.
8.7 Purified water
9.
ASSAY PROCEDURE
9.1 Reconstitute the PTH-MM II Control Level 1 and Level 2 and allow liquid
reagents to equilibrate to room temperature. Mix all reagents gently before
using.
9.2 Set up labeled 12 x 75 mm glass tubes in duplicate according to the Scheme
of the Assay, on the back page.
9.3 Add reagents as follows:
a.
Total count tubes
Set aside until step number 5
b.
Nonspecific binding (NSB)
100 µL of calibrator 0
c.
Calibrator 0
100 µL of Calibrator 0
200 µL of PTH-MM II antiserum
d.
PTH-MM II calibrators (A-F)
100 µL of PTH-MM II calibrator
200 µL of PTH-MM II antiserum
e.
Qualilty control and unknown sera
100 µL of serum
200 µL of PTH-MM II antiserum
9.4 Vortex the tubes gently without foaming and incubate for 15 (±5) minutes at
20-25°C.
* g = (1118 x 10
10:45 AM
) (radius in cm) (rpm)
-8
2
4
Specimens should not be
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