Loop-mediated isothermal amplification reaction primer for detecting streptococcus agalactiae of tilapias

A ring-mediated isothermal, streptococcus lactis technology, applied in the direction of microorganisms, microorganism-based methods, biochemical equipment and methods, etc., to achieve the effect of simple operation, reduced possibility, and significant economic benefits

Inactive Publication Date: 2015-04-29
福建省淡水水产研究所
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At present, no effective drugs have been developed...

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Embodiment 1

[0024] 1. Primer design for LAMP experiments

[0025] Streptococcus agalactiae according to GenBank sip Comparing the gene sequence with the Streptococcus agalactiae isolated from the diseased fishery in our province, select sip The conserved sequence at the front of the gene was analyzed and designed with Primer Explorer IV software, respectively: sip -F3, sip -B3, sip -FIP and sip -BIP, see SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4 for specific sequences.

[0026] 2. strain

[0027] Including 2 strains of Streptococcus agalactiae from tilapia (isolates from diseased tilapia), 8 strains from other Streptococcus species (Streptococcus iniae, Streptococcus dysgalactiae, 2 strains of Streptococcus suis type II, Streptococcus bovis, horse Enterococcus, Streptococcus pyogenes, Streptococcus thermophilus, etc.) and 6 common water strains (Aeromonas hydrophila, Bacillus subtilis, 2 strains of Aeromonas caviae, 2 strains of Escherichia coli).

[0028] 3. B...

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Abstract

The invention relates to a loop-mediated isothermal amplification primer for detecting streptococcus agalactiae of tilapias. The primer is designed according to a conserved sequence of a surface immunogenic gene of the streptococcus agalactiae of the tilapias and comprises sip-F3, sip-B3, sip-FIP and sip-BIP respectively shown by SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4. A loop-mediated isothermal amplification method for detecting the streptococcus agalactiae of the tilapias can be established according to the designed primer. In the method, cross reactions with several common pathogenic bacteria of aquatic animals and several common pathogenic bacteria of streptococcus cannot happen. The detection method provided by the invention does not need expensive instruments, and is short in time, simple in operation and high in sensitivity.

Description

technical field [0001] The invention belongs to the field of aquatic animal bacteriology, in particular to a ring-mediated isothermal amplification reaction primer for detecting tilapia streptococcus agalactiae. Background technique [0002] With the large-scale cultivation of tilapia, the incidence of streptococcosis in tilapia is getting higher and higher. The wide area of ​​the disease and the severity of the damage have become an important obstacle to the development of the tilapia aquaculture industry, causing major damage to the farmers. Economic losses. In the newly revised "List of Types I, II, and III Animal Diseases" by the Ministry of Agriculture in 2008, fish streptococcosis was listed as a type III animal disease, and its main pathogens were Streptococcus agalactiae and Streptococcus iniae. At present, no effective drugs have been developed in China to prevent and control tilapia streptococcosis. Therefore, the establishment of a rapid detection system for til...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/14C12N15/11C12R1/46
CPCC12Q1/6844C12Q1/689
Inventor 郑磊樊海平吴斌曾占壮
Owner 福建省淡水水产研究所
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