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Detection of mycoplasma contamination in veterinary biological product
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Basic data
| Standard ID | GB/T 41699-2022 (GB/T41699-2022) |
| Description (Translated English) | Detection of mycoplasma contamination in veterinary biological product |
| Sector / Industry | National Standard (Recommended) |
| Classification of Chinese Standard | B41 |
| Classification of International Standard | 11.220 |
| Word Count Estimation | 14,168 |
| Date of Issue | 2022-10-14 |
| Date of Implementation | 2023-05-01 |
| Issuing agency(ies) | State Administration for Market Regulation, China National Standardization Administration |
GB/T 41699-2022: Detection of mycoplasma contamination in veterinary biological product
---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.
Detection of mycoplasma contamination in veterinary biological product
ICS 11.220
CCSB41
National Standards of People's Republic of China
Test method for exogenous mycoplasma in veterinary biological products
Published on 2022-10-12
2023-05-01 Implementation
State Administration for Market Regulation
Released by the National Standardization Administration
foreword
This document is in accordance with the provisions of GB/T 1.1-2020 "Guidelines for Standardization Work Part 1.Structure and Drafting Rules of Standardization Documents"
drafted.
Please note that some content of this document may be patented. The issuing agency of this document assumes no responsibility for identifying patents.
This document is proposed and managed by the National Technical Committee on Standardization of Inspection Methods for Key Products of Quality Supervision (SAC/TC374).
This document is drafted by. China Veterinary Drug Administration.
The main drafters of this document. Luo Yufeng, Shen Qingchun, Meng Jie, Zheng Cunzhe, Qu Hongfei, Wang Fang, Liu Bo, Geng Renhao, Wei Jin, Ma Xin, Wang Deli,
Xiao Lu and Chen Dongyang.
Introduction
At present, the mycoplasma testing methods of live veterinary vaccine products in my country have the disadvantages of long testing period, and the testing results are greatly affected by the quality of the culture medium, etc.
question. This document refers to the mycoplasma test methods for biological products included in the current "Chinese Veterinary Pharmacopoeia" and "Chinese Pharmacopoeia", combined with biological information technology.
According to the technical research, a polymerase chain reaction (PCR) test method for mycoplasma detection was developed, and the mycoplasma test culture method was improved. Book
The formulation of the document can standardize the mycoplasma inspection in the production and inspection of veterinary biological products in my country.
Test method for exogenous mycoplasma in veterinary biological products
1 Scope
This document describes the principles, methods and synthesis of the detection of Mycoplasma exogenous in veterinary biological products by polymerase chain reaction (PCR) method and culture method.
Judgment.
This document applies to the inspection of exogenous mycoplasma in veterinary biological products.
2 Normative references
There are no normative references in this document.
3 Terms and Definitions
The following terms and definitions apply to this document.
3.1
mycoplasma
A prokaryotic microorganism that resembles bacteria but has no cell wall, has a diameter of 50nm to 300nm, and can pass through bacterial filters.
Note. Mycoplasma is the simplest microorganism that can grow and reproduce independently on artificial medium.
important microorganisms.
3.2
16SrRNAgene
The corresponding DNA sequence encoding rRNA in bacterial cells.
4 Principles
The deoxyribonucleic acid (DNA) of the sample is separated from each component by physical and chemical methods, using phenol, chloroform (chlorine)
To remove proteins, fats, polysaccharides and other secondary metabolites in the samples, use ethanol to precipitate and wash the nucleic acid samples,
Obtain purified DNA. The 16S rRNA gene sequence of the existing published mycoplasma species was analyzed, and the design was able to detect mycoplasma and
A general primer for steroids, and a polymerase chain reaction (PCR) method for the detection of exogenous mycoplasma in veterinary biological products and cells.
Using mycoplasma culture medium, through the method of separation and culture, artificially conduct veterinary biological products and cells that may contaminate mycoplasma
Culture, and then judge whether there is mycoplasma contamination in the sample by whether there is mycoplasma growth.
5 Mycoplasma test polymerase chain reaction (PCR) method
5.1 Reagents and Materials
Unless otherwise specified, only analytical grade reagents were used.
5.1.1 Balanced phenol. Prepare according to the method of A.1.1 in Appendix A.
5.1.2 Phenol.chloroform.isoamyl alcohol (25.24.1, volume ratio). Prepare according to the method of A.1.
5.1.3 Running buffer (TAE). Prepare according to the method in A.2.
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