GB/T 17999.10-2008 English PDF

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GB/T 17999.10-2008: SPF chicken -- Microbiological surveillance -- Part 10: Indirect immunofluorescent assay for SPF chicken
Status: Valid

Standard ID	USD	BUY PDF	Lead-Days	Standard Title (Description)	Status
GB/T 17999.10-2008	189	Add to Cart	3 days	SPF chicken -- Microbiological surveillance -- Part 10: Indirect immunofluorescent assay for SPF chicken	Valid

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Basic data

Standard ID: GB/T 17999.10-2008 (GB/T17999.10-2008)
Description (Translated English): SPF chicken -- Microbiological surveillance -- Part 10: Indirect immunofluorescent assay for SPF chicken
Sector / Industry: National Standard (Recommended)
Classification of Chinese Standard: B41
Classification of International Standard: 11.220
Word Count Estimation: 8,825
Date of Issue: 2008-12-31
Date of Implementation: 2009-05-01
Older Standard (superseded by this standard): GB/T 17999.9-1999
Regulation (derived from): National Standard Approval Announcement 2008 No.27 (Total No.140)
Issuing agency(ies): General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China, Standardization Administration of the People's Republic of China
Summary: This standard specifies the indirect immunofluorescence test of technical requirements. This section applies to the SPF chickens chicken infectious anemia virus (Chicken Infectious Anaemia Virus, CIAV) to detect antibodies.

GB/T 17999.10-2008: SPF chicken -- Microbiological surveillance -- Part 10: Indirect immunofluorescent assay for SPF chicken

---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.
SPF chicken Microbiological surveillance Part 10. Indirect immunofluorescent assay for SPF chicken ICS 11.220 B41 National Standards of People's Republic of China Replacing GB/T 17999.9-1999 SPF chickens microbiological monitoring Part 10. SPF chickens indirect immunofluorescence assay Posted 2008-12-31 2009-05-01 implementation Administration of Quality Supervision, Inspection and Quarantine of People's Republic of China Standardization Administration of China released

Foreword

GB/T 17999 "SPF chickens microbiological monitoring" is divided into 10 parts. --- The first part 1. SPF chicken microbiological monitoring General; Part --- Article 2. SPF chicken hemagglutination inhibition test; Part --- Section 3. SPF chicken serum neutralization test; Part --- Chapter 4. SPF chicken serum plate agglutination test; Part --- Article 5. SPF chicken agar diffusion test; --- Part 6. SPF chickens ELISA; Part --- Article 7. SPF chicken embryo susceptibility testing; Part --- Article 8. SPF chicken Salmonella testing; --- Part 9. SPF chickens tube agglutination test; Part --- Article 10. SPF chickens indirect immunofluorescence assay. This section GB/T 17999 Part 10. This Part replaces GB/T 17999.9-1999 "SPF chickens indirect immunofluorescence assay." This section compared with the GB/T 17999.9-1999 The main changes are as follows. --- Increasing the normative Appendix A "Preparation of Reagents" and informative Appendix B "cell counting method"; --- A detailed definition of the experimental conditions and the establishment of sample to be tested criteria. Appendix A of this section is normative appendix, Appendix B is an informative annex. This section proposed by the People's Republic of China Ministry of Agriculture. This part of the National Animal Epidemic Prevention Standardization Technical Committee (SAC/TC181) centralized. This section is drafted. Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, China Animal Health and Epidemiology Center, Jinan Spa law Adams Poultry Limited. The main drafters of this section. song even East Liujia Sen, Han Lingxia, satellite Shao, Zhu fruit, single Zhong Fang, Jiang Qian, Secretary Chand, Yu Haibo, Mengqing Wen. This part of the standard replaces the previous editions are. --- GB/T 17999.9-1999. SPF chickens microbiological monitoring Part 10. SPF chickens indirect immunofluorescence assay

1 Scope

This section GB/T 17999 specifies the indirect immunofluorescence test technical requirements. This section applies to SPF chickens chicken infectious anemia virus (ChickenInfectiousAnaemiaVirus, CIAV) antibody Testing. Principle 2 Indirect immunofluorescence test for virus-infected cells as antigen and serum to be tested in the specific antibody binding, and then labeled with a fluorescent dye Anti-antibody binding, the formation of the antigen - antibody - anti-antibody complexes. Fluorescent dye under the action of ultraviolet light or violet light, a visible fluorescence excitation Light, and therefore appear to explain the presence of fluorescent markers, but also reflects the presence of specific antibodies. Often using a known antigen and a fluorescent dye Labeled anti-antibody to detect the corresponding antibodies.

3 Reagents and Equipment

3.1 Reagents 3.1.1 CIAV DELROSE virus strains. 3.1.2 negative and positive sera. 3.1.3 was tested serum. 3.1.4 phosphate buffered saline (PBS) (see Appendix A). 3.1.5 fluorescein isothiocyanate (FITC) labeled anti-chicken IgG (diluted according to the instructions). 3.1.6 buffer or glycerol mounting medium (see Appendix A). 3.1.7 acetone. 3.2 Equipment 3.2.1 antigen sheet. 3.2.2 humidified chamber. 3.2.3 37 ℃ incubator. 3.2.4 fluorescence microscopy. Procedure 4 4.1 Preparation of infected cells With DMEM medium containing 15% fetal bovine serum MDCC-MSB1 cells were cultured at 39 ℃ and 5% carbon dioxide environment. Cells grown to 5 × 106 per milliliter when Ge (cell count see Appendix B), inoculated CIAV, and replaced with a bovine serum-containing DMEM 5% newborn Medium, cultured 36h ~ 48h. 4.2 Preparation of antigen sheet 4.2.1 cytopathic (cell volume increased 2 to 3 times) from about 50% to 75%, the cell cultures were collected to 1500r/min centrifugal 10min, supernatant was discarded, the cell pellet was washed with PBS three times, and the cells were counted, resuspended in PBS cell concentration. 4.2.2 Take 10μL cell suspension (containing about 10 cells 5), was added dropwise in each well of the antigen in the sheet, while normal cells prepared pictures. 4.2.3 drip dry good antigen slices with ice-cold acetone fixed 4 ℃ 10min, stored at -20 ℃, use within a week. ......

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