Burkholderia detection method based on gyrB gene sequence

A technology of Burkholderia and K. spp. is applied in the field of Burkholderia detection based on gyrB gene sequence, which can solve the problems of unfavorable nucleic acid sequencing result accuracy, inconvenient operation, etc.

Pending Publication Date: 2019-12-31
SHANGHAI INST FOR FOOD & DRUG CONTROL
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disclosed UP-1E/APrU primer pair can amplify the gyrB gene fragment, but the length of the amplified product is about 1000bp (longer), which is not conducive to the accuracy of the subsequent nucleic acid sequencin

Method used

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  • Burkholderia detection method based on gyrB gene sequence
  • Burkholderia detection method based on gyrB gene sequence
  • Burkholderia detection method based on gyrB gene sequence

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Embodiment 1

[0041] On the one hand, this embodiment provides specific amplification primers for the gyrB gene, the sequence of which is:

[0042] gyrBF: 5'-CCSACSGACGTGAAGATG-3';

[0043] gyrBR: 5'-TCCACTGCATSGCGACTTC-3'.

[0044]The design process of the primer pair is as follows: using the Burkholderia strain gyrB gene sequence as the screening condition, search the corresponding strain sequence information in the Genbank database, download the fasta file, import the Mega7.0 software for sequence comparison, and obtain the nucleic acid sequence The conserved region and specific region, and primers were designed based on the sequence of the conserved region.

[0045] On the other hand, this embodiment provides a Burkholderia detection method based on the gyrB gene sequence, and the specific steps are as follows:

[0046] 1. Extract the genomic DNA of the experimental strain.

[0047] 2. Nucleic Acid Amplification

[0048] (1) Prepare the amplification system

[0049]

[0050] (2)...

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Abstract

On the one hand, the invention provides a burkholderia detection method based on a gyrB gene sequence. According to the burkholderia detection method, the level of 'species' in genus of burkholderia is identified by amplifying a gyrB gene of the burkholderia, and sequences of amplification primer pairs of the gyrB gene are shown as SEQ ID No.1 to SEQ ID No.2; and on the other hand, the invention provides an integrated detection kit for the level of 'species' in genus of the burkholderia. The kit comprises the primer pairs with the sequences shown as SEQ ID No.1 to SEQ ID No.2. In conclusion, the burkholderia detection method and the integrated detection kit can be used for detecting and identifying burkholderia pollutants in medicine, cosmetics and food safety quality control.

Description

technical field [0001] The invention relates to the field of Burkholderia detection, in particular to a Burkholderia detection method based on the gyrB gene sequence. Background technique [0002] Burkholderia (Burkholderia) is a genus of Burkholderiaceae, Gram-negative bacilli, of which Burkholderia cepacia (Burkholderiacepacia) is the most common, the strain exists in a wide range, can promote biofilm It is a high-risk pollutant in the safety and quality control of pharmaceuticals, cosmetics and food. [0003] "Bergey's Manual of Systematic Bacteriology" (Bergey's Manual of Systematic Bacteriology) is the gold standard for bacterial taxonomy and identification. The genus Burkholderia contains a total of 16 "species" of strain information, among which Burkholderia onion B. cepacia, B. multivorans, B. stabilis, B. vietnamiensis, etc. belong to onion Burkholderia Cepacia Complex (Burkholderia Cepacia Complex, BCC) is currently the focus of attention. BCC is a general term ...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/686C12Q1/04
CPCC12Q1/689C12Q1/686C12Q2535/122C12Q2537/165
Inventor 冯震杨美成秦峰蒋波杨燕李芳肖珊珊
Owner SHANGHAI INST FOR FOOD & DRUG CONTROL
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