Fibroquant kit contains:

1. Thrombin reagent, which is a lyophilized preparation

from bovine source ~50 NIH units per vial.

2. Fibrinogen calibrator, which is a lyophilized preparation of human plasma equivalent to stated amount of

fibrinogen on a mg basis (refer Fibroquant graph paper

supplied with each kit for the value of each lot).

3. Owren’s buffer, ready to use (pH 7.35).

Storage and Stability

1. Store the unopened reagent vials at 2–8°C. Do not

freeze.

2. The shelf-life of the reagents is as per the expiry date

mentioned on the reagent vial labels.

3. Once reconstituted the Fibroquant thrombin reagent

is stable for 6 days when stored at 2-8°C and for 4 hours

at room temperature (20–25°C), provided it is not

contaminated. Extreme care has to be taken to maintain

aseptic precautions while reconstituting, retrieving

and handling reagents to prevent contamination. The

reagent vial must be replaced to 2–8°C immediately

upon retrieving the reagent for the day’s work.

4. The reconstituted Fibroquant fibrinogen calibrator

is stable for 6 hours at 2–8°C and for 2 hours at room

temperature (20–25°C).

Principle

The addition of thrombin coagulates fresh citrated plasma.

The coagulation time is proportional to the fibrinogen

concentration. This allows the estimation of plasma

fibrinogen by functional clotting assay.

Note

1. In vitro diagnostic reagent for laboratory and professional use. Not for medicinal use.

2. The individual reagents contain 0.1% sodium azide as

preservative.

3. Fibroquant thrombin reagent is not from a human

source hence, contamination due to HBsAg, HIV and

HCV is practically excluded.

4. Fibrinogen calibrator provided in the Fibroquant kit is

from a human source, which was tested and found to

be non-reactive for HBsAg, HCV and HIV. However, no

known test methods can assure that infectious agents

are absent. Handle all human products as potentially

infectious.

5. It is very important that absolutely clean and dry

micropipettes be used to aspirate and dispense the

reagent.

6. Avoid exposure of the reagent to elevated temperatures,

direct light and contamination. Immediately

replace the cap after use and store at recommended

temperature.

300 Concise Book of Medical Laboratory Technology: Methods and Interpretations Quality Control

A known normal control should be run in parallel with

each batch of tests. This control may be Tulip plasma

coagulation control Plasmatrol-I or freshly drawn normal

plasma.

Sample Collection and Preparation

No special preparation of the patient is required prior to

sample collection by approved techniques. Withdraw

blood without undue venous stasis and without frothing

into a plastic syringe fitted with a short needle of 19 to

20 SWG. The venipuncture must be a ‘clean’ one and,

if there is any difficulty, take a new syringe and needle

and try another vein. Transfer the blood into tubes,

after detaching the needle from the syringe. Mix nine

parts of freshly collected blood with one part of sodium

citrate (0.109 mol/L, 3.2%). Centrifuge immediately for

15 minutes at 3000 rpm (approximately 2000 g) and

transfer the plasma into a clean test tube. Plasma must be

tested within 3 hours of collection.

Additional Material Required

10 × 75 mm glass test tubes, 0.2 mL and 0.1 mL precision

pipettes, stopwatch, water bath at 37°C, distilled water,

automated, semiautomated/mechanical/optical instrument

if applicable.

Procedure

Bring all the reagents and samples to room temperature

before testing.

Procedure for Fibrinogen Calibration Curve

Preparation

1. The Fibroquant thrombin reagent vial must be

reconstituted exactly with one mL of distilled water;

wait for 5 minutes, do not shake but gently swirl the vial

till the solution attains homogeneity. Further keep the

vial aside for 10 minutes to attain equilibrium. Once

reconstituted it is ready to use for the fibrinogen test.

2. The Fibroquant fibrinogen calibrator vial must be

reconstituted with exactly one mL of distilled water;

wait for 5 minutes, do not shake, gently swirl the vial

till the solution attains homogeneity. Further keep the

vial aside for 10 minutes to attain equilibrium. This is

the fibrinogen calibrator stock solution.

3. Dilute fibrinogen calibrator stock solution with

Owren's buffer as follows:

Test tube no. I II III

Owren's buffer NIL 0.8 mL 0.9 mL

Fibrinogen calibrator 0.2 mL 0.2 mL 0.1 mL

Dilution (calibrator) NIL 1:5 1:10

• Pipette 0.2 mL of each fibrinogen calibrator dilution

into clean test tubes and prewarm for 3 minutes at

37°C.

• Add 0.1 mL of reconstituted thrombin reagent (prewarmed at 37°C for one minute) and simultaneously

start stopwatch.

• Stop the stopwatch at the first appearance of the

fibrin web, as the gel clot begins to form and record

the time in seconds.

• Repeat steps 1 to 3 for a duplicate test one each calibrator dilution.

• Plot the average of the duplicate test values on Tulip

firbrinogen graph paper*.

• Connect the points, which should produce a straight

line.