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NY/T 543-2002: Micro-neutralization test for bovine ephemeral fever
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| Standard ID | Contents [version] | USD | STEP2 | [PDF] delivered in | Standard Title (Description) | Status | PDF |
| NY/T 543-2002 | English | 319 |
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Micro-neutralization test for bovine ephemeral fever
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NY/T 543-2002
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Basic data | Standard ID | NY/T 543-2002 (NY/T543-2002) | | Description (Translated English) | Micro-neutralization test for bovine ephemeral fever | | Sector / Industry | Agriculture Industry Standard (Recommended) | | Classification of Chinese Standard | B41 | | Classification of International Standard | 11.220 | | Word Count Estimation | 10,130 | | Date of Issue | 2002-08-27 | | Date of Implementation | 2002-12-01 | | Summary | This standard specifies the bovine ephemeral fever micro serum neutralization test technologies. This standard applies to the diagnosis of bovine ephemeral fever, immune surveillance and epidemiological investigation. | NY/T 543-2002: Micro-neutralization test for bovine ephemeral fever---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.
Micro-neutralization test for bovine ephemeral fever
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People's Republic of China Agricultural Industry Standard
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Cattle epidemic heat micro-neutralization test method
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Implementation
Published by the Ministry of Agriculture
Foreword
Cattle fever (BEF) is also known as temporary heat, three-day fever, one-day fever, and stiff disease. The disease is found in many countries in Asia, Africa and Oceania.
There have been. The disease is an acute and heat-borne disease caused by the temporary fever virus of the Rhabdoviridae, which is a cyclical epidemic.
During the onset of the disease, it has a serious impact on the milk production, capacity, weight gain, and semen quality of the cattle. It can also cause a small number of sick calves, miscarriages and even
death. Seriously endanger the development of the cattle industry.
This standard is based on China's scientific research results, research results in cooperation with Australia and China's practical experience.
Appendix A, Appendix B and Appendix C of this standard are normative appendices, and Appendix D is an informative appendix.
This standard was proposed by the Ministry of Agriculture, Animal Husbandry and Veterinary Bureau.
This standard is under the jurisdiction of the National Animal Quarantine Standardization Technical Committee.
This standard was drafted. Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences.
The main drafters of this standard. Bai Wenbin, Yan Yuduan.
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Cattle epidemic heat micro-neutralization test method
range
This standard specifies the neutralization test technique for bovine epidemic heat.
This standard applies to the diagnosis, immunological monitoring and epidemiological investigation of bovine epidemic fever.
. Equipment
.. Nutrient solution and 0.05% trypsin EDTA (abbreviated as ATV) solution. See Appendix A for the preparation method.
.. Cells and their preparation, passage, counting methods, see Appendix B.
.. Antigen (indicating toxicity), standard positive serum, and negative serum are provided by the legal unit and used according to the instructions. Antigen toxicity and work resistance
The original (100TCID50) determination method is shown in Appendix C.
.. Microtiter plate. sterile 96-well microtiter plate with lid (referred to as microplate).
.. Micro-sampler. 25μL, 50μL and 100μL single-channel and multi-channel pipette, add plastic dripper (loading 25μL~
100μL, used with the sampler).
.. Carbon dioxide incubator.
. Method of operation
.. Inactivated
The tested serum and the negative and positive serum were inactivated in a 56 ° C water bath for 30 min before use.
.. Preparation
Prepare a working antigen (indicative toxic) method (see Chapter C.1), and dilute the qualified toxicant species with a growth fluid (see Chapter A.1) into
100 TCID50 stewed mL.
.. Qualitative test
... Add 25 μL of growth solution to each well of the microplate.
... In the first row, the serum to be tested is taken up in 2 wells, 25 μL per well, and mixed evenly, which is diluted 2 times.
... Use a multi-channel applicator to transfer 25 μL of liquid from the first well into the second row of wells and mix to form a 4-fold dilution. Then suck from the hole
Discard 25 μL.
... Add 25 μL of working antigen to each well, and mix for 60 min at 37 °C.
... Add 100 μL (30000 cells) of cell suspension to each well. See Section B.3 for cell counting methods.
... The setting of the test control system. Control system settings for each batch of trials. standard positive serum, from column 1 to column 6, each column
4 wells, serially diluted (ie 2 to 64 times), standard negative serum, cell control and virus control, 1 column, 4 wells. Also used in this test
The working antigen was measured again with the test.
... Cover the microplate with a sterile lid and place it in a 37 ° C carbon dioxide (CO2) incubator for 5 days. Record daily from the 3rd day
Cytopathic effect (CPE).
... Result judgment. standard positive serum neutralizing antibody price ≥ 64, standard negative serum, virus control row, CPE appeared in each hole, work
The titration of the antigen content is within the specified range (see Chapter D.1 for the number of pores and the toxic price of CPE), and the normal condition of the cell control well.
Next, 4 times diluted serum was tested, and CPE was found to be negative in both wells; CPE was found in only 1 well, which was judged to be suspicious and should be repeated.
Once the test was performed; if no CPE was produced in both wells, it was judged as positive.
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