Chromogenix Coatest® APC™ Resistance – VS is an APTT-based assay kit for the screening of factor-V-related APC resistance. The high sensitivity and specificity of the test for the factor V:Q506 mutation is obtained by prediluting the sample plasma with an excess of V-DEF Plasma bioreagent. The test design makes it possible to discriminate between heterozygous and homozygous factor V genotypes. It also allows for analysis of plasma from patients on heparin or oral anticoagulant therapy. High discrimination between genotypes with 100% sensitivity for FV:Q506. Reduces need for PCR determination. Applicable to anticoagulant treated patients.

A time- and money-saving alternative to individual genetic testing, Coatest© APC™ Resistance testing is an ideal solution for physicians and labs. Each kit was designed to be easy to use for in-house screening, offering fast results and low costs for both large- and small-volume labs. The most common FV Leiden screening test performed, it offers unmatched sensitivity for the FV:Q506 mutation – close to 100% specificity – and is applicable to patients on heparin or warfarin.

One volume of plasma is prediluted with four volumes of V-DEF Plasma. The dilution is then incubated with the APTT reagent for a standard period of time. Coagulation is triggered by the addition of CaCl2 in the absence and presence of exogenous APC and the time for clot formation is recorded.

Activated Protein C (APC) is a regulator of the coagulation cascade, by specifically inactivating factors Va and VIIIa, in the presence of phospholipids and calcium. In most of the cases (more than 90%), Activated Protein C Resistance (APCR) phenotype is caused by a Factor V gene mutation (Factor V Leiden). The mutation, located on Factor V exon 10 (1691 G –> A), of arginine to glutamine on position 506, renders Factor Va resistant to the cleavage by Activated Protein C. This genetic anomaly can be evidenced with a clotting method performed in the presence or the absence of Activated Protein C.

Reagents and Stability:

1. V-DEF Plasma 2 vials Stabilized, lyophilized human plasma, with a low level of factor V activity, containing the heparin antagonist Polybrene®. Reconstitute with 4.0 mL of NCCLS type II water. Allow to stand for 30 minutes at 20-25°C. Swirl gently before use.

2. CaCl2 2 vials 2 mL of calcium chloride, 0.025 mol/L, in Tris buffer containing 0.5% bovine serum albumin.

3.  APTT reagent 2 vials 4 mL of purified phospholipids with colloidal silica as contact activator. Contains a preservative. Mix thoroughly on a Vortex mixer before use.

4. APC/CaCl2 2 vials Human activated protein C colyophilized with CaCl2. Reconstitute with 2.0 mL of NCCLS type II water. Allow to stand for 30 minutes at 20-25°C. Swirl gently before use.

5. Control Plasma Level 1. 1 vial Lyophilized human plasma. Reconstitute with 1.0 mL of NCCLS type II water. Allow to stand for 30 minutes at 20-25°C. Swirl gently before use.

6. Control Plasma Level 2. 1 vial Lyophilized human plasma. Reconstitute with 1.0 mL of NCCLS type II water. Allow to stand for 30 minutes at 20-25°C. Swirl gently before use.

 

IDEAL FOR:

  • Labs looking to bring testing in-house
  • Labs looking for cost savings
  • Ordering physicians looking for a sensitive assay with rapid turnaround
  • Saves the lab from having to perform genetic test on all patients
  • Economical alternative to genetic testing

Activated Protein C (APC) is a regulator of the coagulation cascade, by specifically inactivating factors Va and VIIIa, in the presence of phospholipids and calcium.

In most of the cases (more than 90%), Activated Protein C Resistance (APCR) phenotype is caused by a Factor V gene mutation (Factor V Leiden). The mutation, located on Factor V exon 10 (1691 G –> A), of arginine to glutamine on position 506, renders Factor Va resistant to the cleavage by Activated Protein C. This genetic anomaly can be evidenced with a clotting method performed in the presence or the absence of Activated Protein C.

Activated Protein C Resistance is tested by using a clotting method performed with or without Activated Protein C.

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