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GB/T 18636-2017Diagnostic techniques for bluetongue (English PDF)

蓝舌病诊断技术

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Issued by

SAMR; SAC

Level / Type

National · Recommended

Issue date

November 1, 2017

Implementation date

May 1, 2018

Scope

GB/T 18636-2017 is the English-translated version of 蓝舌病诊断技术.

GB/T 18636-2017 sets out the diagnostic techniques and test procedures for bluetongue in ruminants, for diagnosis, epidemiological investigation and quarantine.

It covers clinical diagnosis from epidemiology, symptoms and pathological changes; virus isolation from blood, organs and semen; serogroup identification by immunoenzyme staining and antigen-capture ELISA; serotyping by micro-neutralization and plaque inhibition tests; detection of viral nucleic acid by RT-PCR and fluorescent RT-PCR; and antibody detection in serum by agar immunodiffusion and competitive ELISA.

Annexes give the preparation of cell culture fluid, test solutions, primers and probes and agar plates, and the Karber method. Compared with the previous edition, clinical diagnosis and the two RT-PCR methods were added.

It replaces GB/T 18636-2002, was issued on 1 November 2017 and has applied since 1 May 2018.

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Document preview — GB/T 18636-2017

National Standard of the People's Republic of China

ICS: 11.220
Classification: B 41
Replacing: GB/T 18636-2002

Issued by: State Administration for Market Regulation; Standardization Administration of the PRC

Foreword
1 Scope
2 Normative references
3 clinical diagnosis
3.1 Epidemiology
3.2 Clinical symptoms
3.3 pathological changes
3.4 determine the results
4 virus separation
4.1 equipment
4.2 Reagents
4.3 Test Procedure
4.4 virus identification
4.5 results to determine
5 immune enzyme staining
5.1 equipment
5.2 Reagents
5.3 Test Procedure
5.4 test conditions established
5.5 determine the results
6 antigen capture enzyme-linked immunosorbent assay (AC-ELISA)
6.1 Equipment
6.2 Reagents
6.3 Sample
6.4 Experimental Design
6.5 Test Procedure
6.6 Test conditions established
6.7 Determination of results
7 stereotypes and micro-neutralization test
7.1 equipment
7.2 Reagents
7.3 Test Preparation
7.4 Screening Test Procedure
7.5 Test conditions established
7.6 Determination of results
8 plaque and plaque inhibition stereotypes test
8.1 equipment
8.2 Reagents
8.3 Plaque Test Procedure
8.4 Plaque inhibition test procedure
8.5 Determination of Results
9 Reverse Transcription Polymerase Chain Reaction (RT-PCR)
9.1 Equipment
9.2 Reagents
9.3 Test Procedure
9.4 Test conditions established
9.5 Determination of results
10 Fluorescent RT-PCR assay
10.1 Equipment
10.2 Reagents
10.3 Test Procedure
10.4 Threshold Setting
10.5 Test conditions established
10.6 Result Judgment
11 Agar immunodiffusion test (AGID)
11.1 Equipment
11.2 Reagents
11.3 Test Procedure
11.4 result judgment
12 competitive enzyme-linked immunosorbent assay (C-ELISA)
12.1 Equipment
12.2 Reagents
12.3 Test Procedure
12.4 Reading and Suppression Rate Calculation
12.5 conditions for the establishment of the test
12.6 Result Judgment
13 diagnostic results to determine
13.1 Suspected cases
13.2 confirmed cases
13.3 Recessive Infection
Appendix A (Normative) Cell culture fluid preparation
Appendix B (Normative) Preparation of immunoenzyme staining test solution
Appendix C (Normative) Enzyme-linked immunosorbent assay and determination of micro-neutralization test solution
Appendix D (informative) Karber method
Appendix E (Normative) Plaque and plaque inhibition stereotypes test
Appendix F (Normative) RT-PCR test solution and primer preparation
Appendix G (Normative) Synthesis and preparation of fluorescent RT-PCR primers and probes
Appendix H (Normative) Preparation of agar plates

Foreword

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

This standard replaces GB/T 18636-2002 "blue tongue disease diagnostic technology", compared with GB/T 18636-2002, in addition to editorial changes, The main technical changes are as follows.

--- Increased clinical diagnosis;

--- Increased reverse transcriptase polymerase chain reaction (RT-PCR) detection method;

--- Increased fluorescence RT-PCR detection method.

1 Scope

This standard specifies the Bluetongue (Bluetongue, BT) diagnostic techniques and test procedures.

This standard applies to ruminant blue tongue disease diagnosis, epidemiological investigation and quarantine. The methods include clinical diagnosis, pathogen isolation and Identification, pathogenic nucleic acid detection and serological testing, among them.

--- Virus isolation Suitable for isolating Bluetongue virus (BTV) from animal blood, organs and semen;

--- Immunoenzyme staining and antigen capture enzyme-linked immunosorbent assay (AntigenCaptureEnzyme-LinkedImmunosorbent Assay, AC-ELISA) is suitable for the identification of isolated isolates of serogroups;

--- stereotypes and micro-neutralization test and plaque and plaque inhibition stereotyping test for the identification of isolated BTV serotypes;

--- Reverse transcription polymerase chain reaction (Reverse-Transcription Polymerase Chain Reaction, RT-PCR) and fluorescence RT-PCR for the detection of BTV nucleic acids in blood, organs and semen of animals is also suitable for the detection of isolated viruses Serogroup identification

--- Agar immunodiffusion test (AGar) and competitive enzyme-linked immunosorbent assay (CompetitionEnzyme-Linked Immunosorbent Assay, C-ELISA) is suitable for the determination of BTV in serum samples Body detection.

2 Normative references

The following documents for the application of this document is essential. For dated references, only the dated version applies to this edition file. For undated references, the latest edition (including all amendments) applies to this document.

GB/T 6682 analytical laboratory water specifications and test methods

GB 19489 laboratory biosafety common requirements

SN/T 1193 genetic testing laboratory technical requirements

3 Clinical diagnosis

3.1 Epidemiology

3.1.1 Bluetongue disease is caused by the Bluetongue virus of the genus Reoviridae and is transmitted by bites from the civets to ruminants The incidence of non-contact transmission of arbovirus disease.

3.1.2 BTV mainly causes morbidity and mortality in sheep, cattle and goats are often recessive infections, occasional onset and death.

3.1.3 The occurrence and distribution of blue tongue disease is closely related to the library library, mainly in the summer and autumn of the large number of activities in library library, especially in ponds, rivers More common low-lying areas.

3.1.4 Susceptible animals have strong resistance to oral infections, the incidence of animal secretions and excretions within the virus content is very low, not Cause the spread of blue tongue disease.

3.2 Clinical symptoms

3.2.1 incubation period of 3 days to 9 days. Disease early body temperature of sheep is 40.5 °C ~ 42 °C, was reserved hot type, usually lasts 2 days ~ 3 days.

3.2.2 sick sheep lips edema and congestion, there salivation and runny nose and other phenomena. Oral congestion, was purple or blue-purple, soon oral mucosa […]

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This preview omits tables, figures, formulas and parts of the technical clauses. The complete document — 25 pages — is available in the English PDF.

Referenced standards

Normative references

SN/T 1193

Similar standards

GB/T 18088-2026|GB/T 18635-2026|GB/T 18651-2026|GB/T 19168-2026|GB/T 19915-2026|GB/T 22330.1-2026

Editions of GB/T 18636

Edition	Title	Revision	Status
GB/T 18636-2017	Diagnostic techniques for bluetongue	current edition	Current
GB/T 18636-2002	Diagnostic techniques for bluetongue	previous edition	Superseded

This page sells the current edition, GB/T 18636-2017. Earlier editions are listed for reference only.

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