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Whole-genome sequencing method for molecular typing of haemophilus parasuis strains

A whole genome sequencing, Haemophilus suis technology, applied in the direction of microbe-based methods, microbe measurement/inspection, biochemical equipment and methods, etc., can solve the problem of high cost, low resolution and low feasibility of whole genome sequencing and other issues to achieve fast and efficient result feedback

Inactive Publication Date: 2018-09-28
SOUTH CHINA UNIV OF TECH
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Problems solved by technology

However, the poor resolution seriously affects the diagnosis of clinical strains based on housekeeping gene sequencing results
However, the cost of whole genome sequencing is still high at present, and the calculation and analysis methods need to be further improved, and the feasibility for daily monitoring is still low.

Method used

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  • Whole-genome sequencing method for molecular typing of haemophilus parasuis strains

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Embodiment Construction

[0023] The present invention will be further described in detail below with reference to the examples and drawings, but the implementation of the present invention is not limited thereto. Example:

[0024] 1. Extraction of bacterial genomic DNA

[0025] Bacterial genomic DNA was extracted according to the instructions of the DNA extraction kit (spin column type).

[0026] 2. Detection and sequencing

[0027] The extracted bacterial genomic DNA was detected by agarose gel with a concentration of 1.5%. The buffer was lxTAEBuffer and the voltage was 90V for 30 minutes, stained for 30 minutes, and photographed. figure 1 ,Depend on figure 1 It can be seen that 1 to 4 all represent the DNA bands of the HPS1 strain, indicating that the bacterial genome DNA is completely extracted. Genomic DNA was then purified and whole genome sequenced.

[0028] 3. Obtain the housekeeping gene sequence in the database

[0029] Download the sequence of all housekeeping genes (rpoB, atpD, g3pd, ...

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Abstract

The invention discloses a whole-genome sequencing method for molecular typing of haemophilus parasuis strains. According to the whole-genome sequencing method, through a whole-genome sequence, seven house-keeping genes (rpoB, atpD, g3pd, infB, mdh, _6pgd and frdB) in an MLST typing database of the haemophilus parasuis strains are found, and the allele numbers and sequence types of the house-keeping genes are determined; the whole-genome sequencing method comprises the steps of sample pretreatment, genome DNA extraction and quality detecting and sequencing; and by adopting DNAMAN software, thesequencing result is subjected to multi-sequence comparison and analysis, and the strain sequence type (ST) is obtained. Meanwhile it is found that the STs of the three new alleles and the two haemophilus parasuis strains are both new types. The technique process can be used for classification, identification and genetic diversity study of the haemophilus parasuis strains.

Description

technical field [0001] The invention relates to a molecular typing technology for animal pathogenic bacteria, in particular to a method for typing Haemophilus Parasuis based on the multi-locus sequence typing (MLST) technology for whole genome sequencing. Background technique [0002] Haemophilus parasuis strains (HPS) is a short Gram-negative bacterium of Pasteurellaceae, which mainly harms piglets and young pigs. The pathogen is a common bacterium in the upper respiratory tract of pigs. Under certain conditions, it can invade the body and cause serious systemic diseases. Its main features are fibrinous polyserositis, arthritis and meningitis. The resulting disease is also known as "Grager's disease". [0003] At present, Haemophilus parasuis is widespread in our country and even in the whole world, which brings serious economic losses to the pig industry. It is one of the worldwide diseases and has attracted widespread attention. In addition, Haemophilus parasuis is very...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6869C12Q1/689C12Q1/04C12R1/21
CPCC12Q1/6869C12Q1/689C12Q2600/156
Inventor 闫鹤万锈琳李春玲
Owner SOUTH CHINA UNIV OF TECH
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