YY/T 1649.2-2019Test method for interactions of medical devices with platelets - Part 2: In vitro platelet activation products (beta-TG, PF4 and TxB2) assay (English PDF)

Foreword

YY/T 1649 `` Medical Device and Platelet Interaction Test '', including the following.

--- Part 1. In vitro platelet counting method;

--- Part 2. Determination of platelet activation products (beta-TG, PF4 and TxB2) in vitro. This part is the second part of YY/T 1649.

This section is drafted in accordance with the rules given in GB/T 1.1-2009.

Introduction

There are many methods for evaluating platelets and their activation status, but the most commonly used method for evaluating medical devices and materials in contact with blood It is a simple platelet counting method and a method of releasing platelet particulate matter. This part of YY/T 1649 is in vitro platelet activation test Specific test methods can be used as a supplement to the evaluation of the interaction between medical devices/materials and platelets in GB/T 16886.4.

As an important component of blood, platelets play a key role in preventing bleeding. Platelet particulate matter in blood after contact with device/material The increase indicates that platelets are activated, which will directly affect the blood compatibility of medical devices/materials. Described in this part of YY/T 1649 The in vitro assay of platelet activation products can be used to evaluate the potential adhesion and activation of platelets by medical devices/materials.

1 Scope

This part of YY/T 1649 specifies that when performing blood compatibility evaluation of medical devices/materials in vitro, medical devices/materials and blood Platelet Interaction in vitro Platelet Activation Test Method.

This standard applies to the adhesion and activation of platelets on the surface of medical devices/materials. Evaluation of potential.

The test system established in this section applies to human blood. If animal blood is used, it can be tested on the basis of demonstrating its suitability.

2 Normative references

The following documents are essential for the application of this document. For dated references, only the dated version applies to this document. For undated references, the latest version (including all amendments) applies to this document.

GB/T 16886.1 Biological evaluation of medical devices. Part 1. Evaluation and testing during risk management

GB/T 16886.4 Biological evaluation of medical devices. Part 4. Selection of tests for interaction with blood

GB/T 16886.12 Biological evaluation of medical devices. Part 12. Sample preparation and reference materials

3 Terms and definitions

The terms and definitions defined in GB/T 16886.1, GB/T 16886.4, and GB/T 16886.12 apply to this document.

4 Overview

When medical devices/materials come in contact with human blood, the increase in particulate matter in the blood may be caused by medical devices/materials or promote bleeding or Causes of Thrombosis. A significant increase in platelet particulate matter compared to controls indicates that the medical device/material has the potential to activate platelets can. Common platelet particulate matter mainly includes beta-thromboglobulin (beta-TG), platelet factor 4 (PF4), and thromboxane B2 (TxB2) and so on.

5 Test principle (taking beta-TG as an example)

The ELISA kit used the double antibody sandwich method to determine the human beta-TG level in the specimen. Microtiter plates were coated with purified beta-TG antibody to prepare To form a solid phase antibody, beta-TG is sequentially added to the microwells coated with the monoclonal antibody, and then combined with horseradish peroxidase (HRP) labeled beta-TG antibody.

The antibody-antigen-enzyme-labeled antibody complex was formed, and after washing thoroughly, the substrate was added with tetramethylbenzidine (TMB) to develop color. TMB in HRP enzyme It is converted into blue under the catalysis of acid, and finally into yellow under the action of acid. The shade of color is positively correlated with beta-TG in the sample.

The absorbance (OD value) was measured with a microplate reader at a wavelength of 450 nm, and the human beta-TG concentration in the sample was calculated by a standard curve.