GB/T 14926.56-2008Laboratory animal - Method for examination of rabies virus (English PDF)

Foreword

GB/T 14926 "animal" part of a total of 54, different microbes and virus detection methods. Since the implementation of this section instead of GB/T 14926.56-2001 "animal rabies virus detection methods." This part of the main technical differences compared with the GB/T 14926.56-2001 as follows:

a) delete the original standard "2 reference standard" content;

b) the original standard "3 principles" in everything the words "on the pre-coated microplate purified rabies virus antigen in the immune response Performed on a solid support. When the subject serum rabies virus antibodies are present, then the pore walls with an antigen to form an antigen - antibody Complex, and then with anti-dog IgG enzyme conjugate reaction, the final product was measured by enzyme catalyzed substrate after qualitative Detection ";

c) Remove all original standard hemagglutination inhibition test on reagents, methods and results of determination;

d) all of the content of the original standard ELISA test on the changes, including reagents, methods and results criteria, etc.;

e) of the original standard "6 results found" added "immune antibody level pass rate of

6.3 based dog groups. the passing rate of antibody = (Subject animal antibody positive/is the total number of seized animals) × 100%. Ground-level pass rate of antibody dog population reached 70% The penalty for the dog population qualified immunity "content. This part of the National Standardization Technical Committee of experimental animals and focal points. This part of the National Standardization Technical Committee responsible for drafting the experimental animals. The main drafters of this section. TIAN Ke-gong, Chen Xi Zhao, Cao, Wang Chuanbin, Hexiu Min, Tang Hanwen. This part of the standard replaces the previous editions are.

--- GB/T 14926.56-2001. Experimental animal rabies virus detection method

1 Scope

GB/T 14926.56-2008 is the Chinese national standard on laboratory animal - method for examination of rabies virus, in the field of agriculture. The /T suffix marks it as a recommended standard: it is not compulsory by itself, but it becomes binding as soon as a contract, a tender or a customer specification calls it up - which in practice is how most foreign buyers meet it. It was issued on 10 December 2008 by the General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China; Standardization Administration of China, and has been in force since 1 March 2009. Classification: ICS 65.020.30, CCS B44. This page is published from the official record of the standard held by the Chinese standards administration: the identification, the dates, the classification and the issuing body are taken from there. The clause text, the tables and the numeric limits are in the document itself, which is delivered complete in English translation.

GB/T 14926 provisions of this part of the rabies virus (RV) of the detection methods, reagents and the like. This section applies to RV detection dogs. Principle

2 On the plate pre-coated with purified rabies virus antigen, the immune response on a solid carrier. When the subject serum rabies disease When the presence of an antibody drug, the wall of the hole with an antigen to form an antigen - antibody complex, and then reacted with anti-dog IgG enzyme markers, and finally by determining enzyme The product substrates of catalytic after qualitative testing.

3 Reagents and equipment

3.1 Reagents Canine rabies virus IgG antibody test kit (qualitative), comprising.

a) pre-coated microplate rabies virus antigen;

b) rabies virus IgG enzyme label;

c) rabies virus IgG positive control;

d) critical rabies virus IgG control;

e) rabies virus IgG negative control;

f) color reagent A. = sodium hydrogen phosphate - citrate buffer (0.05mol/L, pH5.0) 1000mL, 30% hydrogen peroxide H2O2 (MW34.0) 3.5mL;

g) color reagent B. citric acid 1.052g, disodium ethylenediaminetetraacetate (EDTA-Na2) 0.186g, tetramethylbenzidine (TMB) 0.25g, dimethylsulfoxide (DMSO) 8.0mL, purified water 992mL;

h) Stop Solution. 98% sulfuric acid 27.8mL, purified water 972.2mL;

i) 10-fold concentrated washing solution. PBS (0.1mol/LpH7.2 ~ 7.4) 995mL, Tween -20,5mL;

j) Sample diluent. borate buffer (0.01mol/LpH8.0) 956.0mL, glycerol (MW92.09) 40.0mL, casein (Casein) 10% thimerosal solution 2.0mL, Tween -20,1.0mL1% phenol red solution, 1.0mL. Reagent should be brought to room temperature before use (18 °C ~ 25 °C).

3.2 Equipment Equipment includes.

a) precision pipette 10µL ~ 100µL;

b) a disposable pipette tip;