GB/T 18641-2018Diagnostic method for pseudorabies (English PDF)

Foreword

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

This standard replaces GB/T 18641-2002 "Pseudorabies Diagnostic Technology". Major technical changes compared to GB/T 18641-2002 as follows.

--- Increased blocking ELISA method for detection of pseudorabies virus gB antibody;

--- Added gE-PCR method for detecting pseudorabies virus wild-type virus.

Introduction

Pseudorabies virus (Pseudorabiesvirus, Pr; Aujeszky's disease, AD) is caused by pseudorabies virus (Pseudorabiesvirus, A variety of zoonotic infectious diseases caused by PrV) can harm a variety of animals such as pigs, cattle, sheep, dogs, cats, rabbits, mice, foxes and raccoons.

Worldwide distribution. The disease is the most serious hazard to the pig industry. After the virus infection, pregnant sows have miscarriage, stillbirth and mummified fetuses;

Neurological symptoms occur in piglets at 15 days of age, with a mortality rate of 100%; neurological and respiratory symptoms appear in weaned piglets, with a mortality rate of 10%~ 20%; growth pigs and fattening pigs have respiratory symptoms, slow weight gain, low feed compensation; breeding sows returning to empty and empty, repeated infertility; boar Testicular inflammatory swelling or atrophy, loss of ability to use. In addition to pigs, other animals may be infected with pseudorabies virus, and may have elevated body temperature.

Clinical symptoms such as itching and encephalomyelitis eventually die, but in the form of sporadic.

Conventional serological techniques specified in this standard can be used for the diagnosis of non-immune animals, sero-epidemiological investigations and immunological animal antibody testing.

Among them, the neutralization test has strong specificity and is a common legal method in international trade. It is used for port import and export quarantine; latex agglutination test is simple and fast.

Fast, sensitive, suitable for on-site detection and screening of the disease in the grassroots unit; enzyme-linked immunosorbent assay (ELISA) including whole virus antigen ELISA (PrV-ELISA) and gB-ELISA methods are suitable for laboratory testing of a large number of serum samples. gE- in this standard ELISA differential diagnosis method, which can distinguish between pseudorabies virus gE gene deletion vaccine immunized pigs and naturally infected pigs, so it can be used in pigs Diagnosis and epidemiological investigation of wild-type infections. The three ELISA methods specified in this standard are only used to detect pseudorabies in swine serum.

Viral antibodies are not suitable for detecting pseudorabies virus antibodies in the serum of other animals.

The etiology diagnostic method specified in this standard is used to detect the disease of dead and necropsy animals, aborted fetal tissues (such as tonsils, lungs and Brain tissue, etc.), tonsil in living animals, nasal swabs, and pseudorabies virus in boar semen. Among them, virus isolation and identification are limited to conditions For laboratory use, the polymerase chain reaction is fast and sensitive, allowing simultaneous detection of large numbers of samples, but with careful attention to sample cross-contamination.

Rabbit inoculation tests are carried out in animal rooms with isolation and disinfection conditions.

1 Scope

This standard specifies the serum neutralization test for pseudorabies, latex agglutination test, pseudorabies virus antibody ELISA (PrV-ELISA), Pig pseudorabies gB-ELISA (blocking method) and gE-ELISA (blocking method) and other antibody detection methods and virus isolation and identification, polymerase chain reaction Pathogen detection methods such as inoculation tests with rabbits should be used.

This standard applies to the diagnosis, quarantine, immune antibody evaluation and epidemiological adjustment of pseudorabies in pigs, cattle, sheep, dogs, cats and other susceptible animals. Check and so on.

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

NY/T 678 pig pseudorabies immune enzyme test method

3 Serum neutralization test (fixed virus dilution serum method)