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GB/T 18646-2025 English PDF

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GB/T 18646-2025: Diagnostic techniques for animal brucellosis
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GB/T 18646: Evolution and historical versions

Standard IDContents [version]USDSTEP2[PDF] delivered inStandard Title (Description)StatusPDF
GB/T 18646-2025English1339 Add to Cart 8 days [Need to translate] Diagnostic techniques for animal brucellosis Valid GB/T 18646-2025
GB/T 18646-2018English839 Add to Cart 6 days [Need to translate] Diagnostic techniques for animal brucellosis Valid GB/T 18646-2018
GB/T 18646-2002English479 Add to Cart 3 days [Need to translate] Diagnostic techniques for animal brucellosis Obsolete GB/T 18646-2002

PDF similar to GB/T 18646-2025


Standard similar to GB/T 18646-2025

GB/T 25165   NY/T 1663   GB/T 20365   GB/T 18649   GB/T 18639   

Basic data

Standard ID GB/T 18646-2025 (GB/T18646-2025)
Description (Translated English) Diagnostic techniques for animal brucellosis
Sector / Industry National Standard (Recommended)
Classification of Chinese Standard B41
Classification of International Standard 11.220
Word Count Estimation 66,654
Date of Issue 2025-05-30
Date of Implementation 2025-12-01
Older Standard (superseded by this standard) GB/T 18646-2018
Issuing agency(ies) State Administration for Market Regulation, China National Standardization Administration

GB/T 18646-2025: Diagnostic techniques for animal brucellosis

---This is a DRAFT version for illustration, not a final translation. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.) will be manually/carefully translated upon your order.
National Standard of the People's Republic of China ICS 11.220CCS B 41 Diagnostic technology of animal brucellosis 2025-05-30 Release 2025-12-01 Implementation State Administration for Market Regulation The National Standardization Administration issued Replaces GB/T 18646-2018

Table of contents

Preface...Ⅴ Introduction VI 1 Scope...1 2 Normative references ...1 3 Terms and Definitions 1 4 Abbreviations 1 5 Clinical diagnosis 2 5.1 Popular characteristics 2 5.2 Clinical symptoms 2 5.3 Pathological changes 3 5.4 Result determination 3 6 Sample collection, storage, transportation and processing ...3 6.1 Instruments and Equipment 3 6.2 Reagents and Materials 3 6.3 Sample collection 3 6.4 Sample transportation and storage 4 6.5 Sample processing 4 7 Etiological diagnosis methods 5 7.1 Modified Ziehl⁃Neelsen acid-fast staining...5 7.2 Isolation and identification of Brucella 6 7.3 Real-time fluorescence PCR method 7 7.4 Fluorescence RAA method 9 7.5 Multiplex PCR method...11 8 Immunological diagnostic methods...12 8.1 Standard serum 12 8.2 Red tiger plate agglutination test (RBT) 13 8.3 Tube Agglutination Test (SAT) 14 8.4 Complement fixation test (CFT) 17 8.5 Indirect ELISA antibody detection method (iELISA) ...22 8.6 Competitive ELISA Antibody Detection Method (cELISA) ...24 8.7 Fluorescence Polarization Assay (FPA) 26 8.8 Milk Ring Test (MRT) (for cattle only) 29 8.9 Immunochromatography Assay (ICA) ...30 9 Comprehensive judgment 30 9.1 Determination of animals suspected of Brucella infection 30 9.2 Determination of Brucella Infection in Animals 31 Appendix A (Informative) Applicability of the test method 32 Appendix B (Normative) Preparation of Brucella Culture Medium 33 B.1 Basic culture medium 33 B.2 Serum Glucose Medium 33 B.3 Selection medium 33 Appendix C (Normative) Preparation of Biochemical Reagents 34 C.1 Preparation of Crystal Violet Stock and Working Solutions 34 C.2 Preparation of oxidase test reagents 34 C.3 Urease activity test medium formula and preparation ...34 Appendix D (Normative) Biochemical Identification of Brucella Species and Biotypes 35 Appendix E (Normative) Primer and probe sequences for real-time fluorescence PCR, RAA and multiplex PCR ...37 Appendix F (Normative) Multiplex PCR Solution Preparation and Result Determination 39 F.1 TAE electrophoresis buffer 39 F.2 1.5% agarose gel...39 F.3 Multiplex PCR judgment criteria 39 Appendix G (Normative) Preparation of tube agglutination test (SAT) reagents 40 G.1 Physiological saline containing 0.5% phenol...40 G.2 10% sodium chloride solution containing 0.5% phenol...40 G.3 Preparation of reference turbidimetric tubes 40 Appendix H (Normative) Preparation and titer determination of complement fixation test (CFT) reagents 41 H.1 Preparation of dilution solution 41 H.2 Preparation of Sheep Red Blood Cell Suspension (2.5%) 41 H.3 Hemolysin titer determination 41 H.4 Complement preparation and titer determination 44 H.5 Determination of antigen titer...49 H.6 Preparation of hemolysis standard colorimetric tubes and determination of complement fixation test results ...50 H.7 Dilution and inactivation of serum to be tested 51 Appendix I (Normative) Preparation of ELISA Reagents ...52 I.1 Extraction of coated antigen LPS 52 I.2 Coating solution (0.05 mol/L pH 9.6 carbonate buffer) ...52 I.3 Blocking solution (PBS with 2% BSA and 0.05% Tween-20, pH 7.4) ...52 I.4 Sample diluent (PBS with 1% BSA, pH 7.4) ...52 I.5 Washing solution (PBS with 0.05% Tween-20, pH 7.4) ...52 I.6 Color developing solution 52 I.7 Stop solution (2 mol/L sulfuric acid) 53 Appendix J (Informative) Preparation of Monoclonal Antibodies for Competitive Enzyme-Linked Immunosorbent Assay (cELISA) 54 Appendix K (Normative) Preparation of Fluorescence Polarization Assay (FPA) Antigens and Dilutions 55 K.1 Preparation of labeled antigens 55 K.2 Preparation of sample diluent 55 References ...56

Foreword

This document is in accordance with the provisions of GB/T 1.1-2020 "Guidelines for standardization work Part 1.Structure and drafting rules for standardization documents" Drafting is required. This document replaces GB/T 18646-2018 "Diagnostic techniques for animal brucellosis". Compared with GB/T 18646-2018, except for the structure In addition to adjustments and editorial changes, the main technical changes are as follows. a) The “Scope” has been changed (see Chapter 1, Chapter 1 of the.2018 edition); b) Changed “Clinical Diagnosis” and added content on epidemiology, clinical symptoms and pathological changes (see Chapter 5, 4.1,.2018 edition). 4.2, 4.3); c) Added “real-time fluorescence PCR method” and “fluorescence RAA method” (see 7.3 and 7.4); d) Change “Brucella Bruce-Ladder detection method” to “Multiplex PCR method” (see 7.5, 4.13 of the.2018 version); e) Added the standardization content of RBT, SAT, CFT, iELISA, cELISA and FPA methods in immunological diagnostic methods (see Chapter 8); f) Changed the “Red tiger plate agglutination test (RBT)” (see 8.2, 4.4 of the.2018 edition), “Indirect ELISA antibody detection method (iELISA)” (see 8.5, 4.8 of the.2018 edition), “competitive ELISA antibody detection method (cELISA)” (see 8.6,.2018 4.9 of the.2011 edition); g) Added “Fluorescence polarization assay (FPA)” and “Immunochromatographic assay (ICA)” (see 8.7 and 8.9). This document was proposed by the Ministry of Agriculture and Rural Affairs of the People's Republic of China. This document is under the jurisdiction of the National Technical Committee on Animal Health Standardization (SAC/TC 181). This document was drafted by. China Animal Health and Epidemiology Center, China Veterinary Drug Administration, China Animal Disease Prevention and Control Center Center, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Chinese Center for Disease Control and Prevention, Xinjiang Production and Construction Corps Animal Husbandry and Veterinary Work General Office Liaoning Provincial Animal Disease Prevention and Control Center, Inner Mongolia Autonomous Region Animal Disease Prevention and Control Center, Shanghai Animal Disease Prevention and Control Center Center, Henan Provincial Animal Disease Prevention and Control Center, Chongqing Municipal Animal Disease Prevention and Control Center, Zhejiang Provincial Animal Disease Prevention and Control Center, Qingdao Animal Disease Prevention and Control Center. The main drafters of this document are. Sun Shufang, Li Junping, Shao Weixing, Sun Mingjun, Jiang Hai, Nan Wenlong, Tian Lili, Fan Xiaoxu, Yan Xin, Sun Yu, Hu Sen, Li Yan, Zhu Liangquan, Wang Jianlong, Yan Ruoqian, Zhang Cunrui, Li Yan, Gu Guibo, Zhao Hongjin, Dong Chunxia, Zhao Lingyan, Guo Yu, Sun Xiangxiang, Liu Mengda, Zhang Haobo, Sun Shixiong, and Li Jiaqi. The previous versions of this document and the documents it replaces are as follows. --First published in.2002 as GB/T 18646-2002 and first revised in.2018; --This is the second revision.

Introduction

Brucellosis is a zoonotic infectious disease caused by Brucella infecting animals and humans. The Brucella species reported so far include 6 classical species, 5 newly identified species, and some atypical Brucella species of unidentified types. Classical species include Brucella melitensis, Brucella abortus, and Brucella suis. cella suis), Brucella ovis, Brucella canis, and Brucella sarcoma Brucella neotomae; newly identified species include Brucella ceti, Brucella pinnipedia lis), Brucella microti, Brucella inopinata (other animal hosts have not yet been identified), and Brucella papionis; unidentified species include atypical Brucella isolated from rodents, foxes, and frogs. This document mainly stipulates the three smooth Brucella species, namely, Brucella melitensis, Brucella bovis and Brucella suis, which have the greatest impact on animal husbandry. The diagnostic and detection technology content related to animal brucellosis caused by Bacillus thuringiensis only involves the rough type in pathogen identification and PCR detection method. Detection of Brucella epididymis of sheep and canine species. Brucellosis is a serious threat to animal and human health worldwide. The World Animal Health Organization (WOAH) has included brucellosis in the list of notifiable diseases, and the Ministry of Agriculture and Rural Affairs of my country has issued the "List of Class I, II and III Animal Diseases". The List of Disease Species lists brucellosis as a Category II animal disease, and the National Health Commission lists brucellosis as a Category II animal disease. Personnel engaged in sample collection, packaging and transportation, testing and diagnosis related to brucellosis diagnosis must undergo rigorous training. Conduct experimental activities under the appropriate level of biosafety protection conditions. This document refers to Chapter 3.1.4 Brucellosis (bovine, ovine and porcine species) of the WOAH Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. The diagnostic techniques and methods specified in the "Brucei Infection (2022 Edition)" include clinical diagnosis, etiological diagnosis methods (smear microscopy, isolation and identification) determination, real-time fluorescence PCR method) and immunological diagnostic methods [red tiger plate agglutination test (RBT), tube agglutination test (SAT), complement binding Combined test (CFT), indirect ELISA antibody detection method (iELISA), competitive ELISA antibody detection method (cELISA), fluorescence polarization test (FPA) and whole breast ring test (MRT)]; and according to the development of my country's technology, the mature fluorescent RAA method and immunoassay were added Chromatography (ICA). The issuing organization of this document calls attention to the fact that when declaring that the relevant product complies with this document, it may involve the primer and probe sequences in 7.3 of this document. Listed together with "A Molecular Marker for Distinguishing Brucella A19 Vaccine Strain from Wild Strains" (Authorization Announcement No.. CN113186312B), "Identification of Brucella The use of the related patent "SNP Molecular Markers of Bacillus Vaccine S2 and Wild Strains and Their Applications" (Authorization Announcement No.. CN111793704B). The issuing organization of this document takes no position on the authenticity, validity and scope of the patents involved. The patent holder has promised to the publisher of this document that it agrees to license the patent to any organization free of charge on a fair, reasonable and non-discriminatory basis. Or an individual implements a patent when implementing the national standard. The statement of the patent holder has been filed with the issuing agency of this document. Related information It can be obtained through the following contact information. Patent (Grant Announcement No.. CN113186312B) Holder Name. China Animal Health and Epidemiology Center. Address. No. 369, Nanjing Road, Shibei District, Qingdao City, Shandong Province. Patent (Grant Announcement No.. CN111793704B) Holder Name. Institute of Communicable Disease Prevention and Control, Chinese Center for Disease Control and Prevention. Address. No. 155, Changbai Road, Changping District, Beijing. Please note that in addition to the above two patents, some of the contents of this document may still involve patents. The issuing agency of this document does not assume the responsibility for identifying Patent liability. Diagnostic technology of animal brucellosis

1 Scope

This document describes the clinical, etiological and immunological diagnostic methods for animal brucellosis and provides a comprehensive Meet the judgment requirements. This document is applicable to the monitoring of the freedom of herds and individual animals for brucellosis, implementation of eradication programmes, confirmation of suspected or clinical cases, and diagnosis, population prevalence monitoring, etc. Note. Appendix A lists in detail the selection of detection methods in different fields.

2 Normative references

The contents of the following documents constitute the essential clauses of this document through normative references in this document. For referenced documents without a date, only the version corresponding to that date applies to this document; for referenced documents without a date, the latest version (including all amendments) applies to This document. NY/T 541 Technical Specification for Collection, Storage and Transportation of Veterinary Diagnostic Samples

3 Terms and definitions

There are no terms or definitions that require definition in this document.

4 Abbreviations

The following abbreviations apply to this document. cELISA. Competitive Enzyme Linked Immunosorbent Assay CFT. Complement Fixation Test Ct. Cycle Threshold DNA. Deoxyribonucleic Acid FPA. Fluorescence Polarisation Assay HRP. Horseradish Peroxidase iELISA. Indirect Enzyme Linked Immunosorbent Assay LPS. Lipopolysaccharide OD value. Optical Density PCR. Polymerase Chain Reaction RAA. Recombinase Aided Amplification RBT. Rose Bengal Test SAT. Serum Agglutination Test SDS. Sodium dodecyl sulfate

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