SN/T 1556-2020 English PDFUS$319.00 · In stock
Delivery: <= 4 days. True-PDF full-copy in English will be manually translated and delivered via email. SN/T 1556-2020: (Technical Specifications for Quarantine of Chicken Infectious Rhinitis) Status: Valid SN/T 1556: Historical versions
Basic dataStandard ID: SN/T 1556-2020 (SN/T1556-2020)Description (Translated English): (Technical Specifications for Quarantine of Chicken Infectious Rhinitis) Sector / Industry: Commodity Inspection Standard (Recommended) Classification of Chinese Standard: B41 Classification of International Standard: 65.020.30 Word Count Estimation: 15,198 Date of Issue: 2020-12-30 Date of Implementation: 2021-07-01 Older Standard (superseded by this standard): SN/T 1556-2005 Regulation (derived from): General Administration of Customs Announcement No. 136 [2020] Issuing agency(ies): General Administration of Customs SN/T 1556-2020: (Technical Specifications for Quarantine of Chicken Infectious Rhinitis)---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.Quarantine protocol for chicken infectious coryza The People's Republic of China Entry-Exit Inspection and Quarantine Industry Standards Replace SN/T 1556-2005 Technical Specifications for Quarantine of Chicken Infectious Rhinitis 2020-12-30 release 2021-07-01 implementation Issued by the General Administration of Customs of the People's Republic of China ForewordThis document is drafted in accordance with GB/T 1.1-2020. This document replaces SN/T 1556-2005 "Infectious Rhinitis Agar Immune Diffusion Test Operating Procedures". Compared with SN/T 1556-2005, the main technical changes of this document are as follows. --Added clinical diagnosis methods; --Added pathogen isolation and identification methods; --Added PCR detection method; --Added hemagglutination inhibition test (HI); - Added indirect enzyme-linked immunosorbent test (I-ELISA). This document was proposed and managed by the General Administration of Customs of the People's Republic of China. Drafting organizations of this document. Shenzhen Customs of the People's Republic of China, Shenzhen Institute of Inspection and Quarantine. The main drafters of this document. Tao Hong, Sun Jie, Zhang Caihong, Lin Yanxing, Yang Junxing, Lin Qingyan, Qin Zhifeng, Liu Jianli, Tang Jin Ming Dynasty, Chen Shukun, Liao Lishan, Cao Chenfu, Lu Jianqiang, Hua Qunyi. The previous versions of the documents replaced by this document are as follows. Technical Specifications for Quarantine of Chicken Infectious Rhinitis1 ScopeThis document stipulates the isolation and identification method of infectious rhinitis in chickens, polymerase chain reaction technology, serum plate agglutination test, blood Operation method of coagulation inhibition test, indirect enzyme-linked immunosorbent test, chicken infectious rhinitis agar immunodiffusion test. This document is applicable to the diagnosis and quarantine of infectious rhinitis in chickens.2 Normative referencesThe contents of the following documents constitute the indispensable clauses of this document through normative references in the text. Among them, dated quotations Only the version corresponding to that date is applicable to this document; for undated references, the latest version (including all amendments) is applicable Used in this document. GB/T 6682 Analytical laboratory water specifications and test methods GB/T 18088 Entry and Exit Animal Quarantine Sampling SN/T 2123 Regulations on the collection, transportation and storage of test samples for entry and exit animal quarantine3 Terms and definitionsThere are no terms and definitions that need to be defined in this document.4 Diagnostic technology4.1 Comprehensive clinical diagnosis Refer to Appendix A for the clinical symptoms and pathological changes of infectious rhinitis in chickens. 4.2 Isolation and identification of pathogens 4.2.1 Reagents and materials 4.2.1.1 Sterilized cotton swabs, blood agar plates, sterilized vials, catalase. 4.2.1.2 Staphylococcus epidermidis producing nicotinamide adenine dinucleotide (or coenzyme 1, NAD). 4.2.1.3 5% carbon dioxide (CO2) incubator, 4 ℃ or -20 ℃ ~ -30 ℃ refrigerator. 4.2.2 Sample collection and processing The collection, storage and transportation of samples shall comply with the relevant sampling requirements of GB/T 18088 and SN/T 2123. Collect the blood of diseased chickens aseptically and separate the serum. The serum should be fresh, transparent, non-hemolytic, and non-polluting, and packed in a sterile vial at 4 ℃ Or -20 ℃ ~ -30 ℃ refrigerator storage or refrigerated conditions immediately send for inspection. Aseptically collect the mucus, serous, or secretion of trachea and air sacs in the infraorbital sinus of sick chickens in the acute stage (incubation period 1 d~7 d) It is used for pathogenic diagnosis, and the bacteria in the infraorbital sinus cavity are the purest. 4.2.3 Isolation and identification of bacteria The infraorbital sinus of the suspected chicken was opened aseptically, and the mucus or serum was dipped in a sterile cotton swab. Use a cotton swab to cross the blood agar plate Scribe 5 to 7 lines, and then take NAD-producing Staphylococcus epidermidis and draw a vertical line from the middle of the horizontal line. Place the seeded plate in 5% dioxide Incubate in a carbon incubator at 37 ℃ for 18 h~24 h, and observe the colony characteristics. Take the purified substance of the suspicious colony for catalase test, if there is a large amount Bubbles are positive for catalase. PCR can also be used to identify suspicious colonies, and at the same time, disease materials or suspicious colonies can be taken for animal testing. See Appendix B for the method. 4.2.4 Judgment of results If “satellite-like” dew-like, needle-sized small colonies appear on the blood agar medium, they are close to the NAD-producing epidermal grapes The colony at the coccus line is larger, with a diameter of up to 0.3 mm. The farther away from the NAD-producing Staphylococcus epidermidis line, the smaller the colony will be. Catalase is negative. The result is judged to be positive; if there is no satellite phenomenon, but there are dewdrop-shaped, needle-sized small colonies and catalase is negative, it still needs to be used PCR or animal test identification to avoid missed detection of Avian Bacillus paragallinarum that does not require NAD. If there is no such phenomenon, it is judged as negative. 4.3 PCR detection 4.3.1 Reagents and materials 4.3.1.1 Unless otherwise specified, the reagents used in the test are of analytical grade, and the laboratory water should meet the requirements of GB/T 6682. 4.3.1.2 0.5% Np-40 (ethyl phenyl polyethylene glycol), 0.5% Tween-20, 100 mmol/L pH 8.3 Tris-HCl, 0.2 mg/mL egg White enzyme K, negative and positive controls. 4.3.1.3 Physiological saline, distilled water, 1.5 mL and 0.5 mL small centrifuge tubes, and pipette tips are all sterilized. 4.3.1.4 Thermal cycler, water bath, small centrifuge, micropipette, ice cubes. 4.3.1.5 Agarose, electrophoresis apparatus, horizontal electrophoresis tank, ultraviolet lamp or gel imager. 4.3.1.6 10 mg/mL ethidium bromide, loading buffer, TAE running buffer. 4.3.2 Experimental operation 4.3.2.1 Nucleic acid extraction 4.3.2.1.1 Nucleic acid extraction from tissue samples Open the infraorbital sinus of the suspected chicken aseptically, and dip the mucus or serum into it with a sterile cotton swab and immerse it in 0.8 mL~1.0 mL normal saline. Put the cotton swab on the tube wall as much as possible after placing it in the 1.5 mL centrifuge tube at room temperature for 0.5 h, and then discard it in the waste tank. Centrifuge the centrifuge tube soaked with cotton swabs at 8 000 r/min~10 000 r/min for 10 min, carefully aspirate most of the supernatant, and save 20 µL Around the liquid. Use a pipette to mix the sediment in the centrifuge tube with the remaining liquid evenly, and add 15 µL of 1× PCR buffer (containing 0.5% Np-40, 0.5% Tween-20, 0.2 mg/mL proteinase K and 100 mmol/L pH 8.3 Tris-HCl), press the centrifuge tube cap tightly, and water bath at about 50 ℃ for 1 h, Then 95 ℃ for 10 min, then ice bath for 10 min, PCR amplification or storage at -20 ℃. 4.3.2.1.2 Nucleic acid extraction of colonies Pick the colonies cultured in a 5% carbon dioxide incubator at 37 ℃ for 18 h~24 h, and add them to 0.5 mL small cells containing 10 µL distilled water. In the centrifuge tube, press the tube cap tightly, heat at 95 ℃ for 10 min, then perform PCR amplification after 10 min in ice bath or store at -20 ℃. Use 10 µL of sterile water as a negative control sample, and use 9 µL of sterile water plus 1 µL of a known positive control as a positive control sample. approximately 50 ℃ water bath for 1 h, then 95 ℃ for 10 min, then ice bath for 10 min, PCR amplification or storage at -20 ℃. 4.3.2.1.3 Other nucleic acid extraction methods Equivalent commercial kits can be used for bacterial pathogen nucleic acid extraction. 4.3.2.2 PCR amplification Design PCR detection primers for the hagA gene of Avian paragallinarum. 4.3.2.3 Electrophoresis Prepare 1.2% agarose gel plate. Take 5 µL of PCR product and mix with 2 µL of loading buffer, add it to the agarose gel plate In the hole. Add molecular weight standards. Cover the electrophoresis apparatus, insert the electrodes, and perform electrophoresis at 60 V to 80 V for 30 min to 50 min. Under ultraviolet light Observe the photograph archive; or scan the picture archive with a UV gel imager. Compare the size of PCR fragments with molecular weight standards. 4.3.2.4 Judgment of results The test conditions are established. the negative control has no target band, and the positive control has a specific amplified band at about 510 bp. The test is determined to be established. A band with a molecular weight of 510 bp appears in the test sample, which is the same size as the band that migrates synchronously with the positive control PCR product, and is judged to be a chicken Infectious rhinitis nucleic acid is positive; the test sample does not have any specific band at about 510 bp, which is the same as the negative control, and it is judged as a chicken infectious nose Inflammatory nucleic acid is negative. 4.4 Serological testing 4.4.1 Serum plate agglutination test 4.4.1.1 Material 4.4.1.1.1 Chicken infectious rhinitis plate agglutination test antigen, negative control serum, positive control serum, together with the tested serum before use Return to room temperature. 4.4.1.1.2 Glass slide, 1 micropipette, and sterile tip. 4.4.1.2 Operating procedures 4.4.1.2.1 This test is carried out at room temperature (20 ℃ ~ 25 ℃). 4.4.1.2.2 First write the number on the back of the glass plate, leaving at least 20 mm gap between each sample, and then add 15 µL of antigen on the side of each serial number. 4.4.1.2.3 Add 15 µL of control or test serum according to the serial number on the glass plate, change a tip for each serum sample, and mix the serum with the antigen. The reaction lasts for 3 min to 5 min. Observe the result during this time. Each test has a negative and positive control. 4.4.1.2.4 Observe and record the results on a black background or in a bright place under the lamp. 4.4.1.3 Results judgment and presentation method See Table 1 for the criteria for determining the serum plate agglutination test. Table 1 Judgment criteria for serum plate agglutination test Number reaction performance judgment record symbol qualitative distinction 1 There are obvious frame-like or flaky agglutinates, the liquid is clear and strong positive 2 There are a lot of needle-like particles, the amount is large, and the liquid is clear and positive 3 Needle-like granular, small amount, slightly cloudy and weakly positive 4 The liquid has no particles, uniform and turbid negative-- 4.4.2 Hemagglutination inhibition test 4.4.2.1 Materials 4.4.2.1.1 Chicken infectious rhinitis hemagglutination inhibition test antigen and negative and positive control serum are provided by designated units. 4.4.2.1.2 Normal saline and tested serum. 4.4.2.1.3 Fresh red blood cell suspension and hydroformylated red blood cell suspension, the preparation method sees D.2 and D.3 in Appendix D. 4.4.2.1.4 96-well micro hemagglutination plate (V type), micro shaker, micro pipette and pipette tip. 4.4.2.2 Operating procedures This operation is carried out in the range of 20 ℃~25 ℃, and at the same time, the following operations are performed on the tested serum with A, B, and C antigens respectively. To detect the infection of various types of bacteria. Take a clean hemagglutination plate, and dilute the antigen with physiological saline starting from 5 times to 2 times of incremental dilution and add it to columns 1 to 11 In the holes (that is, the first column is 5×, the second column is 10×, the third column is 20×, and so on to the 11th column), the final liquid volume in each hole is 100 µL, only normal saline is added to the hole in the 12th column as a blood cell control hole. Add 1% of the corresponding red blood cell 100 µL to each well (use fresh A-type antigen) Red blood cells, B and C type antigens use aldehyde erythrocytes), shake and mix thoroughly, and let stand at room temperature for 30 min~60 min (use the blood cells in the control wells) The red blood cells sink completely). Determine the hemagglutination titer of the antigen (HA titer), which is the maximum dilution factor of the antigen that makes the red blood cells fully agglutinated. Absorption of tested serum. Dilute the tested serum 5 times with 10% of the corresponding red blood cells (use fresh red blood cells for type A antigen, B, C Use aldehyde-formed erythrocytes for type antigen), act for 4 h at room temperature or overnight at 4 ℃, fully shake at least 5 times in the middle, and then centrifuge at 1 500 r/min After 10 minutes, the supernatant is taken as the 5-fold diluted serum to be tested. Take a clean blood coagulation plate, start from the first row of holes, and use normal saline to Test serum, negative control serum, and positive control serum, start from 5 times and do 2-fold increasing serial dilutions, measure one serum for each line, and the most for each well. The final liquid volume is 50 µL. Dilute the antigen with normal saline to a concentration of 4 HA titer units (for example, the HA titer is 1.160, then the antigen Dilute to 40 times), add 50 µL to each serum well. Shake well and let stand at room temperature for more than 30 minutes. Add 1% of the corresponding red to each hole Blood cells 100 µL (fresh red blood cells for type A antigens, and aldehyde erythrocytes for type B and C antigens). Shake well, put it at room temperature for 30 min~ ......Tips & Frequently Asked Questions:Question 1: How long will the true-PDF of SN/T 1556-2020_English be delivered?Answer: Upon your order, we will start to translate SN/T 1556-2020_English as soon as possible, and keep you informed of the progress. The lead time is typically 2 ~ 4 working days. The lengthier the document the longer the lead time.Question 2: Can I share the purchased PDF of SN/T 1556-2020_English with my colleagues?Answer: Yes. The purchased PDF of SN/T 1556-2020_English will be deemed to be sold to your employer/organization who actually pays for it, including your colleagues and your employer's intranet.Question 3: Does the price include tax/VAT?Answer: Yes. Our tax invoice, downloaded/delivered in 9 seconds, includes all tax/VAT and complies with 100+ countries' tax regulations (tax exempted in 100+ countries) -- See Avoidance of Double Taxation Agreements (DTAs): List of DTAs signed between Singapore and 100+ countriesQuestion 4: Do you accept my currency other than USD?Answer: Yes. If you need your currency to be printed on the invoice, please write an email to Sales@ChineseStandard.net. In 2 working-hours, we will create a special link for you to pay in any currencies. Otherwise, follow the normal steps: Add to Cart -- Checkout -- Select your currency to pay.Question 5: Should I purchase the latest version SN/T 1556-2020?Answer: Yes. Unless special scenarios such as technical constraints or academic study, you should always prioritize to purchase the latest version SN/T 1556-2020 even if the enforcement date is in future. Complying with the latest version means that, by default, it also complies with all the earlier versions, technically. |
