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Diagnostic techniques for animal anthrax
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Basic data
| Standard ID | GB/T 45101-2024 (GB/T45101-2024) |
| Description (Translated English) | Diagnostic techniques for animal anthrax |
| Sector / Industry | National Standard (Recommended) |
| Classification of Chinese Standard | B41 |
| Classification of International Standard | 11.220 |
| Word Count Estimation | 22,248 |
| Date of Issue | 2024-12-31 |
| Date of Implementation | 2025-07-01 |
| Issuing agency(ies) | State Administration for Market Regulation, China National Standardization Administration |
GB/T 45101-2024: Diagnostic techniques for animal anthrax
---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.
ICS 11.220
CCSB41
National Standard of the People's Republic of China
Diagnostic Techniques for Anthrax in Animals
Released on 2024-12-31
2025-07-01 Implementation
State Administration for Market Regulation
The National Standardization Administration issued
Table of Contents
Preface III
Introduction IV
1 Scope 1
2 Normative references 1
3 Terms and Definitions 1
4 Abbreviations 1
5 Biosafety measures 1
6 Clinical Diagnosis 2
6.1 Popular characteristics 2
6.2 Clinical symptoms 2
6.3 Pathological changes 2
6.4 Result determination 3
7 Sample collection, storage, transportation and processing 3
7.1 Sample collection 3
7.2 Storage and transportation of samples 3
7.3 Sample processing 3
8 Laboratory Diagnosis 4
8.1 Staining microscopy 4
8.2 Isolation and identification 5
8.3 Gamma phage lysis test and penicillin inhibition test 6
8.4 PCR 6
8.5 Real-time PCR 8
8.6 Ring sedimentation test (Ascoli test) 9
9 Comprehensive judgment 10
Appendix A (Normative) Preparation of Reagents 11
A.1 Ammonium oxalate crystal violet staining solution 11
A.2 Gram's iodine solution11
A.3 Sabin yellow counterstain 11
A.4 Alkaline methylene blue staining solution 11
A.5 5%~7% horse or sheep blood agar plate 11
A.6 PLET agar plate 11
A.7 Ordinary nutrient broth 11
A.8 Ordinary nutrient agar plate 11
A.9 0.7% sodium bicarbonate agar plate 11
A.10 50×TAE electrophoresis buffer 12
A.11 2% agarose gel 12
A.12 0.5% phenol saline 12
Appendix B (Informative) Primers and probe sequence information for nucleic acid detection and amplification results Figure 13
B.1 Bacillus anthracis PCR primer 13
B.2 Electrophoresis of PCR amplification products of Bacillus anthracis Figure 13
B.3 Real-time PCR primers and probes 13
B.4 Real-time fluorescence PCR amplification curve of Bacillus anthracis PA gene Figure 14
B.5 Real-time fluorescence PCR amplification curve of Bacillus anthracis CA gene Figure 14
Appendix C (Informative) Target gene sequence of Bacillus anthracis PCR positive control recombinant plasmid 15
C.1 Reference sequence of recombinant plasmid containing protective antigen gene (PA) of Bacillus anthracis 15
C.2 Reference sequence of recombinant plasmid of Bacillus anthracis capsule gene (CA)15
Appendix D (Informative) Ring Sedimentation Test Results Figure 16
Reference 17
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.
Please note that some of the contents of this document may involve patents. The issuing organization of this document does not assume the responsibility for identifying patents.
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/TC181).
This document was drafted by. Beijing Animal Husbandry and Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, Military Veterinary Research Institute of the Military Medical Research Institute of the Academy of Military Sciences
Institute, China Veterinary Drug Administration, Beijing Animal Disease Prevention and Control Center.
The main drafters of this document are. Ding Jiabo, Qin Tong, Zhu Lingwei, Jiang Hui, Guo Xuejun, Zhang Yinghui, Fan Xuezheng, Peng Xiaowei, Feng Yu, Wang Lin,
Cheng Rujia.
Introduction
Anthrax is an acute, febrile, septicemic zoonotic disease caused by Bacillus anthracis.
The disease is widely distributed throughout the world. The disease mainly occurs among livestock, with herbivores such as cattle, sheep, and horses being the most susceptible, followed by omnivores.
The disease is mainly transmitted through contact with animals infected with anthrax or contaminated animal products or environmental infection. Animal infection with anthrax is mainly divided into the most acute type, acute
There are three clinical types. acute, subacute and chronic. Each type has a high mortality rate and poses a serious threat to human and animal health. World Organization for Animal Health
(WOAH) lists it as a reportable animal disease, and my country lists it as a Category II animal disease.
This document was prepared with reference to Chapter 3.1.1 of the WOAH Manual of Diagnostic Tests and Vaccines for Terrestrial Animals (2023 Edition).
technology, and combined with the new research results of related technologies in my country.
Diagnostic Techniques for Anthrax in Animals
1 Scope
This document describes the clinical diagnosis, staining and microscopy, isolation and identification, gamma phage lysis test, penicillin inhibition test,
Laboratory diagnostic methods such as PCR, Real-time PCR, and ring precipitation test.
This document applies to the diagnosis, detection, quarantine, surveillance and epidemiological investigation of anthrax in animals.
2 Normative references
The contents of the following documents constitute the essential clauses of this document through normative references in this text, among which the dated references
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.
GB 19489 General requirements for laboratory biosafety
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.
Ct value. Cycle Threshold
FAM.6-Carboxy-Fluorescein
NFQ-MGB. Non-Fluorescent Quencher-Minor Groove Binder
PLET. Polymyxin B-Lysozyme-EDTA-Thalousace-
tateAgar)
TAE. Tris-Acetate-EDTA Buffer
5 Biosafety measures
It is strictly forbidden to perform autopsies on animals suspected of having anthrax or on animals that have been infected with anthrax under non-biosafety conditions.
Necessary protective measures should be taken to prevent infection of personnel and contamination of the environment. Laboratories engaged in anthrax diagnosis should follow the
Provisions are made for risk assessment and risk control.
...
