T/SDAA 0045-2021 Active Group standards

T/SDAA 0045-2021: Detection of Multi-killing Pasteurella by PCR Method

T/SDAA 0045-2021

Publish Date: 2021-11-30 Implement Date: 2021-12-31 For services related to genuine standard inquiry, procurement, translation, and other related services in China, please Contact Us

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Basic Information

Standard Code: T/SDAA 0045-2021

Standard Type: Group standards

Standard Status: Active

is_force_gb: no

CCS Name: Health

ICS Name: Animal husbandry and breeding

Publish Date: 2021-11-30

Implement Date: 2021-12-31

Scope

Main Technical Content: Detection of Multiple-virulent Pasteurella multocida by PCR 1 Scope This document specifies the PCR detection method for Pasteurella multocida in pigs. This method is applicable to the detection of Pasteurella multocida in pigs. 2 Normative Reference Documents The content of the following documents is an essential part of this document through normative reference. Among them, the referenced documents with a specified date apply only to the version corresponding to that date; the latest version (including all amendment sheets) of the unspecified-date referenced documents applies to this document. - NY/T 541: Technical Specifications for the Collection, Storage, and Transportation of Veterinary Diagnostic Samples - NY/T 564-2016: Diagnostic Techniques for Pasteurella multocida Disease in Pigs 3 Principle A pair of specific primers are designed based on the gene sequence of Pasteurella multocida, and the DNA nucleic acid of Pasteurella multocida is used as the template to amplify the specific DNA fragment. The amplified DNA fragment is electrophoresed on an agarose gel, and the result is judged by the size of the DNA band. 4 Reagents or Materials Unless otherwise specified, all reagents used are analytical grade. 4.1 Water: GB/T6682, Grade I. 4.2 5% Sheep Fresh Blood Agar Plate 4.3 Gram Stain Solution 4.4 2× Taq Master Mix 4.5 DNA Marker DL 2000 4.6 Agarose 4.7 TAE Buffer 4.8 4S Green Plus Non-Toxic Nucleic Acid Dye T/SDAA 0045-2021 5 Instrumentation and Equipment 5.1 Biosafety Cabinet: Equipped with an ULPA ultra-high efficiency air filter. 5.2 High-speed Freeze Centrifuge: Rotation speed not less than 12,000 rpm/min. 5.3 PCR Machine: Maximum temperature and cooling rate of 4℃/s, temperature range 4℃~99℃. 5.4 Gel Imaging System: Image resolution not less than 5 million pixels, sensitivity capable of detecting less than 20pg EB-stained double-stranded DNA. 5.5 UV Transilluminator: Transilluminati