GB/T 18935-2018 English PDF

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GB/T 18935-2018: Diagnostic techniques for foot and mouth disease
Status: Valid

GB/T 18935: Historical versions

Standard ID	USD	BUY PDF	Lead-Days	Standard Title (Description)	Status
GB/T 18935-2018	559	Add to Cart	5 days	Diagnostic techniques for foot and mouth disease	Valid
GB/T 18935-2003	679	Add to Cart	3 days	Diagnostic techniques for foot-and-mouth disease	Obsolete

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Basic data

Standard ID: GB/T 18935-2018 (GB/T18935-2018)
Description (Translated English): Diagnostic techniques for foot and mouth disease
Sector / Industry: National Standard (Recommended)
Classification of Chinese Standard: B41
Classification of International Standard: 11.220
Word Count Estimation: 28,237
Date of Issue: 2018-09-17
Date of Implementation: 2019-04-01
Older Standard (superseded by this standard): GB/T 18935-2003
Regulation (derived from): National Standard Announcement No. 11 of 2018
Issuing agency(ies): State Administration for Market Regulation, China National Standardization Administration

GB/T 18935-2018: Diagnostic techniques for foot and mouth disease

---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.
Diagnostic techniques for foot and mouth disease ICS 11.220 B41 National Standards of People's Republic of China Replace GB/T 18935-2003 Foot and mouth disease diagnosis technology Published on.2018-09-17 Implementation of.2019-04-01 State Market Supervisory Administration China National Standardization Administration issued

Content

Foreword V Introduction VI 1 Scope 1 2 Normative references 1 3 Abbreviations 1 4 clinical diagnosis 1 4.1 susceptible animals 1 4.2 Clinical symptoms 2 4.3 Pathological changes 2 4.4 Result determination 2 5 Laboratory Diagnostic Sample Collection 2 5.1 Equipment 2 5.2 Reagent 2 5.3 Sample Collection 2 5.4 Sample Processing 3 6 virus separation 3 6.1 Equipment 3 6.2 Reagent 4 6.3 Test animals and cells 4 6.4 Test procedure 4 6.5 Virus Identification 5 6.6 Result determination 5 7 stereotyped enzyme-linked immunosorbent assay (scheduled ELISA) 5 7.1 Equipment 5 7.2 Reagent 5 7.3 Test procedure 6 7.4 Test establishment conditions 6 7.5 Result determination 6 8 Multiple Reverse Transcription-Polymerase Chain Reaction (Multiple RT-PCR) 6 8.1 Equipment 6 8.2 Primer 7 8.3 Reagent 7 8.4 Sample Preparation 8 8.5 Test procedure 8 8.6 Test establishment conditions 9 8.7 Determination of results 9 9 stereotyped reverse transcription-polymerase chain reaction (sequencing RT-PCR) 9 9.1 Equipment 9 9.2 Primer 9 9.3 Reagent 9 9.4 Sample Preparation 9 9.5 Test procedure 9 9.6 Test establishment conditions 10 9.7 Result determination 10 10 Virus VP1 gene sequence analysis 10 10.1 Equipment 10 10.2 Primer 10 10.3 Reagent 10 10.4 Sample Preparation 10 10.5 Test procedure 10 10.6 Test establishment conditions 11 10.7 Results Determination and Analysis 11 11 Fluorescence quantitative reverse transcription polymerase chain reaction (fluorescence quantitative RT-PCR) 11 11.1 Equipment 11 11.2 Primers and Probes 11 11.3 Reagent 11 11.4 Sample Preparation 12 11.5 Test procedure 12 11.6 Test establishment conditions 12 11.7 Result determination 12 12 Virus Neutralization Test (VN) 12 12.1 Equipment 12 12.2 Reagent 12 12.3 Test procedure 13 12.4 Test establishment conditions 13 12.5 Determination of results 13 13 Liquid phase blocking enzyme-linked immunosorbent assay (LPB-ELISA) 14 13.1 Equipment 14 13.2 Reagent 14 13.3 Control serum 14 13.4 Test procedure 14 13.5 Test establishment conditions 14 13.6 Result determination 14 14 Solid phase competition enzyme-linked immunosorbent assay (SPC-ELISA) 15 14.1 Equipment 15 14.2 Reagent 15 14.3 Test procedure 15 14.4 Test establishment conditions 15 14.5 Determination of results 15 15 Non-structural protein 3ABC antibody indirect enzyme-linked immunosorbent assay (3ABC-I-ELISA) 15 15.1 Equipment 15 15.2 Reagent 16 15.3 Test procedure 16 15.4 Test establishment conditions 16 15.5 Calculation and Judgment of Results 16 16 non-structural protein 3ABC antibody blocking enzyme-linked immunosorbent assay (3ABC-B-ELISA) 16 16.1 Equipment 16 16.2 Reagent 16 16.3 Test procedure 17 16.4 validity of test results 17 16.5 Determination of results 17 17 Comprehensive judgment 17 Appendix A (Normative) Sample preservation solution and cell culture solution 18 Appendix B (Normative) Preparation of solutions for enzyme-linked immunosorbent assays 20 Appendix C (Normative) Preparation of liquids for nucleic acid detection 22

Foreword

This standard was drafted in accordance with the rules given in GB/T 1.1-2009. This standard replaces GB/T 18935-2003 "Diagnostic Techniques for Foot and Mouth Disease". This standard is compared with GB/T 18935-2003, except for editorial In addition to the changes, the main technical changes are as follows. --- Increased clinical diagnosis; --- Removed the micro-complement binding assay; --- Cancel the part of the virus testing test, the relevant content is specified in other parts; --- Removed the virus infection-related (VIA) antigen agar gel immunodiffusion test (VIA-AGID), increased non-structural protein 3ABC antibody indirect enzyme-linked immunosorbent assay (3ABC-I-ELISA) and non-structural protein 3ABC antibody block enzyme-linked immunosorbent assay Adsorption test (3ABC-B-ELISA) to replace VIA-AGID; --- Increased stereotyped reverse transcription-polymerase chain reaction; --- Increased sequence analysis of the viral VP1 gene; --- Increased fluorescence quantitative reverse transcription-polymerase chain reaction. This standard was proposed by the Ministry of Agriculture and Rural Affairs of the People's Republic This standard is under the jurisdiction of the National Animal Health Standardization Technical Committee (SAC/TC181). This standard was drafted. Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences. The main drafters of this standard. Liu Xiangtao, Zhang Qiang, Guo Jianhong, He Jijun, Liu Zaixin, Lu Zengjun, Bao Huifang, Chang Huizhen, Tian Hong, Zheng Haixue, Shang Youjun, Ma Junwu, Wu Guohua, Takino, Lin Mi, Ma Weimin, Lu Yongqian, Zhu Caizhu. The previous versions of the standards replaced by this standard are. ---GB/T 18935-2003.

Introduction

Foot and Mouth Disease (FMD) is caused by Foot and Mouth Disease (FMDV) An acute, potent, and contact infectious disease caused by hoofed animals. Foot-and-mouth disease can cause huge economic losses and social impact, world animals The Health Organization (OIE) lists foot-and-mouth disease as an animal-borne disease that must be reported. China's foot-and-mouth disease is a class of animal diseases. The source of foot-and-mouth disease is mainly latent infection and clinically ill animals. Susceptible animals can pass through the respiratory tract, digestive tract, reproductive tract and wounds Oral infection of the virus, usually by direct or indirect contact (droplets, etc.), or by human or dog, fly, cockroach, bird and other animal media, or by car Vehicles, appliances, etc. are spread by pollutants. If the climate is right, the virus can spread long distances with the wind. Infected animal exhaled, saliva, feces, Urine, milk, semen, meat and by-products can be poisoned. After infection with ruminants such as cattle and sheep, the virus can continue to be poisoned in the esophagus-throat. The foot-and-mouth disease virus is classified into the picornavirus family (Picornaviridae), the foot-and-mouth disease virus genus, and has seven serotypes, namely O, A, Asia1, C, SAT1, SAT2 and SAT3, there is no cross-immunoprotective response between the serotypes, and the immune control is equivalent to 7 kinds of no The same epidemic, serotype identification is the first problem to be solved by immune prevention and control. Samples suitable for the diagnosis of foot-and-mouth disease are unruptured or newly broken water Bubbles and blister fluids. In the absence of blister and blisters, blood can be collected and/or ruminant food can be collected using an esophageal cup. A sample of tract-pharyngeal secretions, also present in these samples. In the absence of a tissue sample, detection of specific antibodies is also used for diagnosis. Virus isolation of tissue or liquid samples to detect the presence of foot-and-mouth disease virus antigens or nucleic acids can make a positive diagnosis. If sample Insufficient or undefined results, by reverse transcriptase polymerase chain reaction (RT-PCR), and/or by sensitive cell culture or suckling mice The nucleic acid or live virus which may be present in the sample is proliferated and then detected by enzyme-linked immunosorbent assay (ELISA) or RT-PCR. In serological diagnosis, if a specific antibody is detected in an unvaccinated animal, a positive diagnosis can be made. This method is mild for mild cases. Cases with vesicular epithelial samples are not very useful. Detection of certain non-structural protein antibodies to foot-and-mouth disease virus can be used as Evidence of proviral infection. For the identification of foot-and-mouth disease virus type, there are stereotyped enzyme-linked immunosorbent assay (scheduled ELISA), stereotyped Transcription-polymerase chain reaction (typing RT-PCR), a method for the diagnosis of foot-and-mouth disease virus antigens and nucleic acids with multiple reverse transcription-polymerase Chain reaction (multiplex RT-PCR), viral VP1 gene sequence analysis and fluorescence quantitative reverse transcription-polymerase chain reaction (fluorescence quantitative RT- PCR), a method for detecting foot-and-mouth disease virus structural protein antibodies, virus neutralization test (VN), liquid phase blocking enzyme-linked immunosorbent assay (LPB-ELISA) and solid phase competition enzyme-linked immunosorbent assay (SPC-ELISA) for the detection of non-structural protein antibodies to foot-and-mouth disease virus Non-structural protein (NSP) 3ABC antibody indirect enzyme-linked immunosorbent assay (3ABC-I-ELISA) and non-structural protein (NSP) The 3ABC antibody blocked the enzyme-linked immunosorbent assay (3ABC-B-ELISA). The revision of this standard refers to the OIE "Handbook of Diagnostic Tests and Vaccine Standards for Terrestrial Animals" (2017 version), combined with the relevant technology research in China. Research and development of new results, technically consistent with international advanced technology. Foot and mouth disease diagnosis technology

1 Scope

This standard specifies the technical requirements for clinical diagnosis and laboratory diagnosis of Foot and Mouth Disease (FMD). This standard is applicable to the diagnosis of foot-and-mouth disease of cloven-hoofed animals such as pigs, cattle and sheep and other susceptible animals.

2 Normative references

The following documents are indispensable for the application of this document. For dated references, only dated versions apply to this article. Pieces. For undated references, the latest edition (including all amendments) applies to this document. GB 19489 General requirements for laboratory biosafety

3 Abbreviations

The following abbreviations apply to this document. CPE. Cytopathic effect DNA. Deoxyribonucleic acid ELISA. Enzyme linked immunosorbent assay FMD. Foot and Mouth Disease (Footandmouthdisease) FMDV. Foot and Mouth Disease Virus (Footandmouthdiseasevirus) HRP. Horseradish peroxidase (Horseradish peroxidase) ID50. half of the infectious dose (Medianinfectivedose) LPB. Liquid phase block (Liquidphaseblock) MEM. Minimum essential medium (Minimumessentialmedium) NSP. Nonstructural protein OP solution. esophageal-pharyngeal fluids (Oesophageal-pharyngealfluids) OPD. O-Phenylenediamine PBS. Phosphate buffered saline buffer (Phosphatebufferedsalinebuffer) RNA. Ribonucleic acid RT-PCR. Reverse Transcription-polymerase chain reaction (Reversetranscription-polymerase chain reaction) SPC. Solid phase competition (Solidphasecompetition) TCID50. half of the cell infection (Mediantissuecultureinfectivedose) TMB. tetramethylbenzidine (3,3',5,5'-tetramethyl-benzidine) VN. Virus Neutralization Test (Virusneutralisationtest)

4 clinical diagnosis

4.1 susceptible animals Artiodactyls, including bovines (bovine, zebu, buffalo, yak), sheep, goats, and all wild ruminants and pigs The foot-and-mouth disease virus is susceptible, and the camelid is less susceptible. Equine animals are not infected with foot and mouth disease. ......

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