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Diagnostic techniques for avian leukosis
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Basic data
| Standard ID | GB/T 26436-2025 (GB/T26436-2025) |
| Description (Translated English) | Diagnostic techniques for avian leukosis |
| Sector / Industry | National Standard (Recommended) |
| Classification of Chinese Standard | B41 |
| Classification of International Standard | 11.220 |
| Word Count Estimation | 26,229 |
| Date of Issue | 2025-01-24 |
| Date of Implementation | 2025-08-01 |
| Issuing agency(ies) | State Administration for Market Regulation, China National Standardization Administration |
GB/T 26436-2025: Diagnostic techniques for avian leukosis
---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
Avian Leukosis Diagnostic Technology
2025-01-24 Release
2025-08-01 Implementation
State Administration for Market Regulation
The National Standardization Administration issued
Replace GB/T 26436-2010
Table of Contents
Preface Ⅲ
Introduction Ⅳ
1 Scope...1
2 Normative references ...1
3 Terms and Definitions 1
4 Abbreviations 1
5 Laboratory Biosafety Measures...2
6 Clinical diagnosis 2
6.1 Epidemiology 2
6.2 Clinical symptoms 2
6.3 Pathological changes 3
6.4 Differential diagnosis 3
6.5 Clinical judgment 3
7 Sample collection and processing ...4
7.1 Sample Collection 4
7.2 Sample processing 4
8 Laboratory diagnosis...4
8.1 Virus isolation and identification 4
8.2 Identification of viral subgroups 7
8.3 Real-time RT-PCR amplification of ALV-J...7
8.4 Serological diagnosis 7
9 Comprehensive judgment 8
9.1 Suspected...8
9.2 Confirmation 8
Appendix A (Normative) Preparation of solutions related to virus isolation 9
A.1 0.01 mol/L PBS (pH 7.4)...9
A.2 50% glycerol-PBS storage solution (pH 7.4)...9
A.3 Complete cell nutrient solution and maintenance solution 9
Appendix B (Informative) Preparation of CEF (C/E strain) 10
Appendix C (Informative) Preparation of anti-ALV single factor chicken serum 11
Appendix D (Normative) Preparation of solutions related to p27 antigen ELISA detection ...12
D.1 Coating solution 12
D.2 Washing liquid 12
D.3 Sample diluent 12
D.4 Substrate solution 12
D.5 Stop Solution 12
Appendix E (Informative) ALV Subpopulation Identification Procedure 13
E.1 Cloning vector and host bacteria 13
E.2 Preparation of viral templates 13
E.3 Amplification of the envelope protein env gene 13
E.4 Cloning of PCR products 14
E.5 Extraction and identification of plasmid DNA 15
E.6 Determination of clone sequence 15
Appendix F (Informative) Real-time RT-PCR Amplification of ALV-J...17
F.1 Primer and probe sequences 17
F.2 Sample collection and pretreatment 17
F.3 Nucleic Acid Extraction 17
F.4 Preparation of amplification reagents 17
F.5 Sample addition test 18
F.6 Reaction Conditions Setting 18
F.7 Fluorescence Setting ...18
F.8 Analysis condition setting and result judgment 18
Appendix G (Informative) Composition and Use of ALV-J One-Step Real-time RT-PCR Detection Kit ...19
G.1 Kit Composition...19
G.2 Description 19
G.3 Precautions when using...19
Appendix H (Informative) Preparation of ALV-infected Cells for Antibody Detection by IFA Method 20
References ...21
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 26436-2010 "Diagnostic technology for avian leukosis". Compared with GB/T 26436-2010, in addition to structural adjustments and
In addition to editorial changes, the main technical changes are as follows.
-- Added "Abbreviations" (see Chapter 4);
--Added the ALV⁃p27 antigen immune colloidal gold test paper card detection method (see 8.1.4.3);
--The criteria for confirming a diagnosis based on antibody test results have been changed (see 8.4.5, 4.3.2.3 of the.2010 edition).
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/TC 181).
This document was drafted by. Shandong Agricultural University, Gongbei Customs Technical Center of the People's Republic of China, South China Agricultural University, Yangzhou University,
Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lingyu Biotech (Beijing) Pharmaceutical Technology Co., Ltd., China Agricultural University, Pulike Biotech
Cheng Co., Ltd., China Animal Disease Prevention and Control Center.
The main drafters of this document are. Zhao Peng, Cui Zhizhong, Sun Shuhong, Yang Su, Sha Caihua, Liao Ming, Cao Weisheng, Qin Aijian, Qian Kun, Wang Yixin,
Gao Yulong, Chen Xizhao, Su Jingliang, Tian Kegong, Wu Qingmin, and Wang Chuanbin.
The previous versions of this document and the documents it replaces are as follows.
--First published in.2010 as GB/T 26436-2010;
--This is the first revision.
Introduction
Avian leukosis (AL) is a multidrug-resistant disease in poultry caused by infection with avian leukosis virus (ALV).
A general term for various neoplastic diseases, which mainly cause malignant tumors and subclinical infections in chickens, leading to growth retardation, decreased egg production and immune suppression in chickens.
It is an immunosuppressive disease that can be transmitted vertically through breeding eggs. my country classifies it as a Class III animal epidemic.
ALV belongs to the genus Alpharetrovirus, subfamily Orthoretrovirinae, family Retroviridae, and is currently classified according to the characteristics of its gp85 protein.
ALV is divided into 11 subgroups. A, B, C, D, E, F, G, H, I, J and K. ALV is divided into two major types. endogenous ALV and exogenous ALV.
Endogenous ALV refers to ALV that is integrated into the host cell chromosome genome and can be vertically transmitted through the chromosome.
Incomplete fragments of the viral genome do not produce infectious virions, but may be complete viral genomes and thus be able to produce infectious virions
However, these viruses are usually of low or no pathogenicity, mainly E subgroup ALV. Exogenous ALV is different from endogenous ALV.
Correspondingly, it refers to ALVs that are not transmitted to the next generation through the host cell chromosome, including A, B, C, D, J and K subgroups.
ALV is an exogenous virus. The interference of endogenous ALV is the first thing to be excluded when conducting laboratory tests, especially nucleic acid tests.
factor.
The disease has no obvious seasonal or regional characteristics and is usually caused by raising chicks whose seed sources are not purified of ALV, as well as by
The disease may be caused by mixing chickens with infected chickens for a period of time or using biological products contaminated with exogenous ALV.
The diagnostic technology content of this document includes clinical diagnosis, etiological diagnosis and serological diagnosis methods.
It is based on the World Health Organization (WOAH)'s Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, and combines it with my country's new research results on related technologies.
Avian Leukosis Diagnostic Technology
1 Scope
This document describes the clinical diagnosis of avian leukosis, sample collection and processing, as well as virus isolation and identification, identification of virus subgroups,
Laboratory diagnostic methods such as real-time RT-PCR, ALV-J amplification, and serological diagnosis.
This document applies to the diagnosis, detection, quarantine, monitoring and epidemiological investigation of avian leukosis in chicken flocks.
2 Normative references
The contents of the following documents constitute the essential clauses of this document through normative references in this document, 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.
document.
GB/T 6682 Specifications and test methods for water used in analytical laboratories
GB/T 18088 Quarantine sampling for entry and exit of animals
GB/T 18643 Diagnostic techniques for Marek's disease in chickens
GB 19489 General requirements for laboratory biosafety
NY/T 541 Technical Specification for Collection, Storage and Transportation of Veterinary Diagnostic Samples
NY/T 1247 Diagnostic techniques for avian reticuloendotheliosis
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.
AL. Avian leukosis
ALV. Avian leukosis virus
CPE. Cytopathic effect
Ct value. Cycle threshold
CEF.Chick embryo fibroblast
DEPC. Diethyl pyrocarbonate
DMEM. Dulbecco's modification of Eagle's medium
dNTP. Deoxyribonucleoside triphosphate
EDTA. Ethylene diamine tetraacetic acid
ELISA. Enzyme-linked immunosorbent assay
IFA. Indirect immunofluorescence assay
MDV. Marek's disease virus
...
