YY/T 1500-2016Pyrogen test for medical devices—Monocyte-activation test—Human whole blood ELISA method (English PDF)

Foreword

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

This standard is based on the European Pharmacopoeia 7.0 monocyte activation test based on the combination of medical device products to develop.

Introduction

Pyrogenic detection is an important item in the safety evaluation of medical devices. Medical devices Pyrogenic reactions are mainly divided into endotoxin-mediated heat The original reaction and non-endotoxin-mediated pyrogenic reaction.

The pyrogen is divided into exogenous pyrogen and internal pyrogen. Common pyrogen is mostly outside the pyrogen, outside the pyrogen into the body can be activated after the body (IL-1beta), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNFalpha), interleukin- And so on. Within the pyrogen can be directly through the blood - brain barrier in the body temperature adjustment center so that the body temperature adjustment point on the move, the body heat increased, Heat dissipation, causing heat.

Monocyte activation test (MAT) is used to activate human monocytes or mononuclear cells to release such as proinflammatory cytokines, IL-1beta, IL-6 and TNF-alpha, and the like. And these cytokines can cause fever disease. Therefore, the MAT can detect the pyrogen present in the test sample. This guideline is given in Appendix A for guidelines for monocyte activation.

In 2009, the European Pharmacopoeia included in vitro pyrogen examination, the US FDA also received five kinds of human immune cells based on the injection of drug fever In vitro test method. These methods use human whole blood, component blood or cell lines as a cell source, to detect the contact with the test substance after incubation IL-1beta, IL-6 and TNF-alpha and other internal pyrogen. International Organization for Standardization Technical Committee (ISO /TC194) 16 Working Group A document It is stated that the detection of IL-6 and IL-1beta has a higher signal-to-noise ratio and is a good cytokine pyrogenic marker. This standard is given in Appendix B An example of human whole blood IL-1beta method for thermoanometry of medical devices.

1 Scope

This standard gives a test method for the determination of pyrogen in medical devices/materials by means of human whole blood ELISA.

2 Normative reference documents

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

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

3 Overview

Medical device/material after contact with human whole blood, the medical device/material contained in the pyrogen-activated blood in the mononuclear cells, activated Mononuclear cells can produce endogenous pyrogen, such as IL-1beta, IL-6 and TNFalpha. The intrahepatic pyrogen in the whole blood was measured by ELISA And the response of the mononuclear cells to the test product and the response to the endotoxin standard, indirectly reflecting the medical device/material The amount of pyrogenic substance contained.

4 Full blood source and quality requirements

5 Utensils

Use a confirmed process to remove all glassware and other heat-resistant devices in a dry-hot oven. Usually used for the time and temperature