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NY/T 680-2003: Enzyme-linked immunosorbent assay for avian leukosis virus p27 antigen
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| NY/T 680-2003 | English | 209 |
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Enzyme-linked immunosorbent assay for avian leukosis virus p27 antigen
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NY/T 680-2003
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Basic data | Standard ID | NY/T 680-2003 (NY/T680-2003) | | Description (Translated English) | Enzyme-linked immunosorbent assay for avian leukosis virus p27 antigen | | Sector / Industry | Agriculture Industry Standard (Recommended) | | Classification of Chinese Standard | B41 | | Classification of International Standard | 11.220 | | Word Count Estimation | 6,630 | | Date of Issue | 2003-07-30 | | Date of Implementation | 2003-10-01 | | Summary | This standard specifies the avian leukosis virus (ALV) p27 antigen ELISA method. This standard applies to detect the egg white and egg embryo tissue cultures of ALVp27 protein antigens. | NY/T 680-2003: Enzyme-linked immunosorbent assay for avian leukosis virus p27 antigen---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.
Enzyme-linked immunosorbent assay for avian leukosis virus p27 antigen
ICS 11.220
B41
People's Republic of China Agricultural Industry Standard
Enzyme linked immunosorbent assay
Released on July 30,.2003
Implementation of.2003-10-01
Published by the Ministry of Agriculture
Foreword
Appendix A of this standard is a normative appendix.
This standard is proposed and managed by the Animal Husbandry and Veterinary Bureau of the Ministry of Agriculture.
This standard was drafted. Veterinary Diagnostic Center of the Ministry of Agriculture.
The main drafters of this standard. Chen Xiwei, Su Jingliang, Wang Hongwei, Tian Kegong, Wu Qingmin, Wang Chuanbin.
Enzyme linked immunosorbent assay
1 Scope
This standard specifies the avian leukosis virus (ALV) p27 antigen enzyme-linked immunosorbent assay.
This standard is applicable to the detection of ALVp27 protein antigen in egg white and chicken embryo tissue cultures.
2.1 Test materials
2.1.1 Reagents
2.1.1.1 Anti-p27 protein antibody. Prepared by immunizing rabbits with purified avian myeloblastosis virus p27 protein.
2.1.1.2 Positive antigen. egg white containing chicken leukemia/sarcoma virus-specific p27 protein antigen.
2.1.1.3 Negative antigen. SPF egg white without chicken leukemia/sarcoma virus-specific p27 protein antigen.
2.1.1.4 Enzyme conjugate. Horseradish peroxidase-labeled anti-p27 protein antibody.
2.1.1.5 Phosphate buffer (PBS). See section A for formulation. Chapter 1.
2.1.1.6 Coating liquid. See Appendix A for preparation. Chapter 2.
2.1.1.7 Washing liquid. See Appendix A for preparation. Chapter 3.
2.1.1.8 Sample diluent. See Appendix A for preparation. Chapter 4.
2.1.1.9 Substrate solution. See Appendix A for preparation. Chapter 6.
2.1.1.10 Stop solution. See Appendix A for preparation. Chapter 7.
2.1.2 Equipment
a) enzyme-linked detector;
b) polystyrene board;
c) micro-sampler, capacity 50μL ~.200μL;
d) 37 ° C constant temperature incubator.
2.1.3 Sample
2.1.3.1 Egg sample. Place the small end of the egg to be inspected upwards, break the eggshell, aseptically absorb the egg white with a pipette, and compact it in a small sterilization.
Inside the bottle.
2.1.3.2 Cell culture. Pipette the cell culture with a pipette, in a sterile vial, store at 4 ° C or -30 ° C or send immediately
Check. The samples to be inspected shall be numbered uniformly before the test, and no dilution will be made during the test.
2.2 Method of operation
2.2.1 The anti-p27 protein antibody was diluted 4000-fold with a coating solution, and 100 μL per well was added to a 96-well polystyrene plate at 4 ° C overnight.
2.2.2 Remove the coated polystyrene board, discard the liquid, rinse each well with 250 μL of washing solution for 3 min each time, and dry.
2.2.3 Add 150 μL of 1% gelatin blocking solution to each well and react at 37 ° C for 60 min.
2.2.4 Repeat 2.2.2.
2.2.5 Add 100 μL standard negative control to the A1 and A2 wells, and add 100 μL standard positive control to each of the A3 and A4 wells.
2.2.6 Add the sample to be tested to each other hole, add two holes to each sample to be tested, 100 μL per well, and react at 37 ° C for 60 min.
2.2.7 Repeat 2.2.2.
2.2.8 Add 100 μL of working concentration of horseradish peroxidase-labeled anti-p27 protein antibody to each well and react at 37 ° C for 60 min.
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