GB/T 18637-2018Technical specifications for diagnosis of bovine viral diarrhea / mucosal disease (English PDF)

Foreword

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

This standard replaces GB/T 18637-2002 "Blood virus diarrhea/mucosal disease diagnosis technology", in addition to editorial changes, the main technical changes The following is as follows.

--- Added "clinical diagnosis" method (see Chapter 4);

--- Added "real-time fluorescent RT-PCR detection" (see 5.4).

Introduction

Bovine Viral Diarrhea/Mucosal Disease (BVD/MD, BVD for short) is caused by bovine disease Toxic diarrhea/mucosal disease virus (BovineViralDiarrhea/Mucosal Disease Virus, BVD/MDV, referred to as BVDV) infection Bovine-induced fever, mucosal erosion ulcers, leukopenia, diarrhea, cough, and miscarriage of pregnant cows or the production of malformed fetuses An infectious disease. BVDV is a taxonomic genus of the genus Flaviviridae and Pestivirus.

The species, currently identified by the International Classification of Viruses, BVDV has two genotypes (or species), namely BVDV-1 and BVDV-2. it They are associated with the same family of swine fever virus (ClassicalSwineFeverVirus, CSFV) and Border Disease Virus (BorderDiseaseVirus, There is a structural and antigenic association between BDV) and a partial cross-reaction in serology. The disease caused by BVDV-1 is globally distributed. More common, and the BVDV-2 caused by the disease is rare. Both genotypes of virus can produce cells based on their cultured cells.

The lesion is divided into two biotypes, Cytopathic Type (CP) and Non-cytopathic (Non-cytopathic). Type, NCP). NCP strains are biotypes that cause all types of infections and pathologies, while CP strains are mainly found in mucosal cases.

At present, the disease is widespread in the world. In 1983, China first isolated BVDV. In recent years, pathogen surveys have shown China's cattle, sheep, deer, pigs, etc. are infected with BVDV to varying degrees, which has seriously affected the healthy development of animal husbandry in China.

The methods described in this standard are commonly used in the diagnosis of BVD in China in recent years, including clinical diagnosis and pathological diagnosis. Broken and serological diagnostic techniques. Serological diagnostic techniques are virus neutralization assays. Pathogenic diagnostic techniques include virus isolation and identification and real-time fluorescence Real RT-PCR (Real-time RT-PCR) detection method, the Real-time RT-PCR detection method in this standard has BVDV specificity Sexuality, capable of simultaneously detecting BVDV-1 and BVDV-2, and specifically distinguishing between BVDV and CSFV.

Combined with China's animal epidemic prevention laws, regulations and policies, this standard can effectively diagnose and monitor the occurrence of BVD/MD in China. popular.

1 Scope

This standard specifies the clinical diagnosis of epidemiology, clinical symptoms, pathological changes of bovine viral diarrhea/mucosal disease, and sample collection and Technical requirements and operational specifications for laboratory diagnostics such as isolation, identification and identification, Real-time RT-PCR and virus neutralization tests.

This standard is applicable to the detection of type 1 and type 2 bovine viral diarrhea/mucosal disease virus, nucleic acid and antibody levels in different samples.

2 Normative references

The following documents are indispensable for the application of this document. For dated references, only dated versions apply to this document. For undated references, the latest edition (including all amendments) applies to this document.

GB/T 6682 Analytical laboratory water specifications and test methods

GB/T 19438.1-2004 Avian influenza virus universal fluorescent RT-PCR detection method

GB 19489 General requirements for laboratory biosafety

GB/T 27401 Laboratory Quality Control Regulations Animal Quarantine

3 Abbreviations

The following abbreviations apply to this document.

BT. Bovine Turbinatecel

BVD/MD. Bovine Viral Diarrhea/Mucosal Disease

BVD/MDV. Bovine Viral Diarrhea/Mucosal Disease Virus

CPE. Cytopathic Effect (CytopathicEffect)

CP. Cytopathic type

cRNA. Complement RNA (ComplementRNA)

Ct(Cp). the number of cycles experienced by the amount of fluorescent signal in each reaction tube reaching a set threshold (CycleThreshold, or CrossingPoint)

DEPC. DiethylPyrocarbonate

FAM. Carboxyfluorescein

FBS. fetal bovine serum (FetalBovineSerum)

FITC. Fluorescein Isothiocyanate

MDBK. Bovine kidney secondary cells (Madin-DarbyBovineKidneycel)

MEM. Basic Medium (MinimumEssentialMedium)

NCP. Non-cytopathic type

PBS. Phosphate Buffer (PhosphateBufferSaline)

PBST. Tween Phosphate Buffer (PhosphateBufferSolutionwithTween20)

PI. Persistent Infection

Real-time RT-PCR. Real-time Reverse Transcription Polymerase Chain Reaction (Real-time Reverse Transcription Polymerase ChainReaction)