SN/T 5106-2019 in English
VALIDDNA barcoding identification of Dengue vector-mosquitoes
- Issued on:2019-09-03
- Implemented on:2020-03-01
- File Format:PDF
- Delivery:Via email within 1~3 business days
$156.00
| Standard No: | SN/T 5106-2019 |
| Document status: | VALID |
| Title in English: | DNA barcoding identification of Dengue vector-mosquitoes |
| Title in Chinese: | 登革热媒介蚊类DNA条形码鉴定方法 |
| Language: | English |
| File Format: | Electronic (PDF) |
| Delivery: | Via email within 1~3 business days |
| Issued on: | 2019-09-03 |
| Implemented on: | 2020-03-01 |
| Professional Classification: | SN-Import&Export Inspection |
| Related Keywords: | core identification process key
precise identification traditional morphological identification standard sequence specific standard sequence |
| Related Topics: | dna
Trace DNA bar code dna+ standard dna Forward and reverse primers dna standard standard dna dengue Mosquito instrument dna/ mosquito mosquitoes dna barcode identification slats dna dna qualitative method dna+tm value how to identify dna Lipid Identification fluorescence identification dna laboratory dna barcode identification DNA barcoding molecular identification screening Dengue fever regulations sy/t 5106-2019 |
Introduction
Technical background and significance of the standard
This standard aims at the precise identification of the main dengue fever vectors, Aedes albopictus and Aedes aegypti, and establishes a standardized process for DNA barcoding based on the mitochondrial COI gene. It was issued and implemented by the General Administration of Customs in 2019, solving the technical bottleneck of traditional morphological identification in the identification of damaged specimens.
Analysis of the core identification process
| Key steps | Technical parameters | Quality control points |
|---|---|---|
| Specimen processing | Unilateral tissue is taken from adults/larvae/pupa and preserved in 90% ethanol | Voucher specimen preparation complies with SN/T1876 |
| DNA extraction | Microtissue lysis method or commercial kit | OD260/280 ratio 1.7-2.0 |
| PCR amplification | Primers LCO1490/HCO2198, annealed at 46℃ | Set positive and negative controls |
Technical key points
COI gene selection basis
Mitochondrial cytochrome c oxidase subunit I (COI) gene has:
1) High inter-species variability (＞2%) and intra-species conservation
2) High universal primer coverage
3) 750bp fragment contains sufficient phylogenetic information
Sequence comparison judgment rules
Use NCBI BLAST tool for double sequence comparison:
- Sequences with similarity ≥98% to the standard sequence are judged to be of the same species
- The primer region (first 20bp and last 25bp) needs to be excluded
Suggestions for the implementation of the standard
- Laboratory hierarchical management: The front and back regions of PCR are strictly separated, in line with the second-level requirements of biosafety in GB19489
- Quality control system: Each batch of testing must include a standard (KY825745/KY825746) control
- Handling of difficult specimens: For samples with a similarity of 97-98%, it is recommended to add nuclear gene marker verification
Technology evolution analysis
Compared with the SN/T4278-2015 version of the operating procedures, this standard:
1) Added a Aedes aegypti specific standard sequence
2) Optimized the annealing temperature to 46℃ to improve specificity
3) Clarify the minimum sampling volume requirements for residual limb tissue
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