T/NAASS 057-2023 Active Group standards

T/NAASS 057-2023 Niu4 type herpesvirus indirect enzyme-linked immunosorbent assay (IELISA) antibody detection technical specification

Publish Date: 2023-03-22 Implement Date: 2023-04-25 For services related to genuine standard inquiry, procurement, translation, and other related services in China, please Contact Us

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Basic Information

Standard Code: T/NAASS 057-2023

Standard Type: Group standards

Standard Status: Active

is_force_gb: no

CCS Name: -

ICS Name: Animal husbandry and breeding

Publish Date: 2023-03-22

Implement Date: 2023-04-25

Scope

Scope: This document specifies the terms and definitions of the antibody detection of Bovine Herpesvirus Type 4 (BoHV-4) by indirect enzyme-linked immunosorbent assay (iELISA), and the application of this method in the detection, diagnosis, and epidemiological investigation of the disease in bovine serum, plasma, and milk samples. Main Technical Content: 3. Terms and Definitions The following terms and definitions apply to this document: - Bovine Herpesvirus Type 4 (BoHV-4) - Indirect Enzyme-Linked Immunosorbent Assay (iELISA) The iELISA method first binds the antigen to the ELISA plate, followed by two detection steps: first, adding the detection antibody to specifically bind with the antigen; then, adding the enzyme-labeled secondary antibody for detection and using a substrate for color development to quantify the antigen based on the color reaction. 4. Operational Procedures for iELISA - Preparation of Experimental Environment, Instruments, and Equipment - Preparation of Washing Solutions - Preparation of Dilution Solutions - Preparation of Tracer Working Solutions All reagents shall be prepared according to the instructions in the kit. The biosafety conditions of the experimental environment shall comply with GB/T 19489, and the distilled water used for dilution shall meet the requirements of GB/T 6682. Samples can be used directly for detection, including fresh or frozen serum, plasma, and milk samples. **Milk Sample Preparation**: - Centrifuge at 4000 rpm/min for 20 minutes. - Use a Pasteur pipette to extract the middle layer liquid, taking care not to touch the lower layer of cell sediment. - Add undiluted skim milk directly to the microplate. **Operational Procedures**: - Restore all reagents to 21°C ± 3°C before use and gently shake or rotate to mix them evenly. - **4.3.1 Preparation**: Remove the microplate and mark the sample position on the operation record sheet. If only a portion of the microplate is us