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GB/T 38165-2019: Detection of circulating cell-free DNA concentration in human peripheral blood - Real-time PCR method based on Alu sequence
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| Standard ID | Contents [version] | USD | STEP2 | [PDF] delivered in | Standard Title (Description) | Status | PDF |
| GB/T 38165-2019 | English | 199 |
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Detection of circulating cell-free DNA concentration in human peripheral blood - Real-time PCR method based on Alu sequence
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GB/T 38165-2019
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Basic data | Standard ID | GB/T 38165-2019 (GB/T38165-2019) | | Description (Translated English) | Detection of circulating cell-free DNA concentration in human peripheral blood - Real-time PCR method based on Alu sequence | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | A40 | | Classification of International Standard | 07.080 | | Word Count Estimation | 10,134 | | Date of Issue | 2019-10-18 | | Date of Implementation | 2019-10-18 | | Issuing agency(ies) | State Administration for Market Regulation, China National Standardization Administration | GB/T 38165-2019: Detection of circulating cell-free DNA concentration in human peripheral blood - Real-time PCR method based on Alu sequence
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Detection of circulating cell-free DNA concentration in human peripheral blood--Real-time PCR method based on Alu sequence
ICS 07.080
A40
National Standards of People's Republic of China
Detection of circulating free DNA concentration in human peripheral blood
Real-time fluorescence PCR based on Alu sequence
Published on.2019-10-18
2019-10-18 implementation
State market supervision and administration
China National Standardization Administration issued
Foreword
This standard was drafted in accordance with the rules given in GB/T 1.1-2009.
This standard is proposed and managed by the National Biochemical Testing Standardization Technical Committee (SAC/TC387).
This standard was drafted. Chengdu University of Traditional Chinese Medicine, Sichuan Cancer Hospital, Shanghai Institute of Metrology and Testing Technology.
The main drafters of this standard. He Yang, Xiao Hongtao, Song Yi, Li Yan, Wen Yanli, Leng Ping, Liu Gang, Li Dongmei.
Detection of circulating free DNA concentration in human peripheral blood
Real-time fluorescence PCR based on Alu sequence
1 Scope
This standard specifies the real-time fluorescent polymerase chain of circulating free DNA (DNA) based on Alu sequence in human peripheral blood.
Reaction (PCR) quantitative detection method.
This standard applies to the quantitative detection of Alu sequences in circulating free DNA samples.
2 Normative references
The following documents are indispensable for the application of this document. For dated references, only dated versions apply to this article.
Pieces. For undated references, the latest edition (including all amendments) applies to this document.
GB/T 6682 Analytical laboratory water specifications and test methods
GB 19489 General requirements for laboratory biosafety
3 Terms and definitions
The following terms and definitions apply to this document.
3.1
Circulating free DNA circulatingcel-free DNA; cfDNA
A short fragment of DNA in the extracellular free state derived from apoptosis and necrosis in the blood.
3.2
Real-time fluorescent PCR real-time PCR
Adding a fluorophore to the PCR reaction system, monitoring the entire PCR amplification process in real time through the accumulation of fluorescent signals, and achieving the initial
Quantitative and qualitative analysis of the template.
3.3
Ct value cyclethreshold
The number of cycles experienced by the fluorescent signal in each reaction tube reaching a set threshold.
3.4
Alu sequence Alusequence
A moderately repeating sequence scattered throughout the primate genome.
Note. Alu sequences have 70% to 98% homology, accounting for 5% to 10% of the human genome sequence, and each element is approximately 300 bp in length.
4 Principle
The extracted peripheral blood circulating free DNA is used as a detection target sample, and the qualified human genomic DNA quantitative standard substance is
Standards, using real-time fluorescent PCR technology, design specific primers to amplify fragments of 115 bp and 219 bp in Alu sequences, respectively.
In the paragraph, the logarithm of the DNA concentration of the standard starting template is the abscissa and the Ct value is the ordinate, and a standard curve is prepared. According to the standard curve equation
The Alu sequence copy number in the sample was calculated and the results are expressed in copies/μL.
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