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Primer set and kit for detecting serotype of streptococcus parauberis

A primer set and serotype technology, applied in biochemical equipment and methods, microorganisms, recombinant DNA technology, etc., can solve the problems of difficulty in changing the serotype of Streptococcus parauberis from fish and long cycle

Active Publication Date: 2019-10-22
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the most commonly used method for serotype detection and identification is serum agglutination test, but this method requires a long cycle, and most research institutes and laboratories do not have antiserum for serotype, so the traditional method is used to identify fish Difficulty in changing the serotype of Streptococcus parauberis

Method used

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  • Primer set and kit for detecting serotype of streptococcus parauberis
  • Primer set and kit for detecting serotype of streptococcus parauberis
  • Primer set and kit for detecting serotype of streptococcus parauberis

Examples

Experimental program
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Effect test

Embodiment 1

[0043]Type identification of serotypes Ia, Ib / Ic and II of Streptococcus parauberis from fish

[0044] The experimental research object is a strain of three different serotypes Ia, Ib / Ic and II of Streptococcus parauberis derived from fish. In addition, strains of Streptococcus parauberis derived from bovine and other common fish pathogenic bacteria are used to detect the specificity of the detection method. sex.

[0045] The above two pairs of primers are added to the detection system at the same time, and the system used for PCR amplification preferably includes: 2 μL of genomic DNA, 2 μL of 10 μM forward and reverse primers (two pairs), 1 μL of 10 mM dNTPS, 0.5 μL of Taq DNA polymerase, 10 μL for each 50 μL of PCR amplification. ×Taqpolymerase buffer 5μL and dd H 2 O 33.5 μL. In the present invention, the PCR amplification program includes: 98°C for 3 minutes; 98°C for 10s, 55°C for 30s, 72°C for 1min, 30 cycles; 72°C for 7min.

[0046] There are three combinations of pr...

Embodiment 2

[0048] Specific detection of 6 strains of serotype Ia streptococcus parauberis derived from fish

[0049] The two pairs of primers added in the detection system have three combinations, namely Parauberis-F2 / Parauberis-R2, Parauberis-F1 / Parauberis-R1 and Parauberis-F1 / Parauberis-R2. Streptococcus uberis strains have all amplified the expected bands, there are three places, the lengths are respectively: 183bp, 428bp and 763bp, this result is consistent with type Ia in Example 1, so it can be used for the detection of fish-derived Streptococcus parauberis Specific detection of serotype Ia strains. The result is as figure 2 shown.

Embodiment 3

[0051] Specific detection of 7 strains of serum Ib / Ic fish-derived Streptococcus parauberis

[0052] As in the detection system of the above-mentioned embodiment 1, the 7 strains of serum Ib / Ic type fish-derived Streptococcus parauberis strains used in the experiment can all amplify the expected band of 428bp size, such as image 3 shown. This result is consistent with the detection result of Ib / Ic type in Example 1, and thus can be used for the specific detection of fish-derived Streptococcus parauberis serotype Ib / Ic strains.

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Abstract

The invention provides a primer set and kit for detecting the serotype of streptococcus parauberis, and belongs to the technical field of fish disease detection. The primer set comprises a Parauberis1primer pair and a Parauberis2 primer pair. The primer set can accurately, specifically, rapidly and sensitively distinguish the serotype of the fish-derived streptococcus parauberis.

Description

technical field [0001] The invention belongs to the technical field of fish disease detection, in particular to a primer set and a kit for detecting the serotype of Streptococcus parauberis. Background technique [0002] Streptococcus parauberis is an important aquatic animal pathogen, which was first identified as a fish pathogen in Spanish cultured turbot in 1996, and then the disease spread in Europe. In North America, the fungus has been reported to cause infection of wild striped bass. In recent years, this pathogen has been the main cause of disease outbreaks of cultured flounder in Asia, Korea and Japan. The infected fish showed symptoms such as eyeball protrusion, hyperemia, muscle hemorrhage, and abdominal distension. Cause acute mass death and cause significant economic losses. At the same time, the latest report of sepsis in cultured freshwater ornamental fish caused by this bacterium was reported in Israel. S. parauberis can be classified into 5 serophenotypes ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/04C12N15/11C12R1/46
CPCC12Q1/689
Inventor 涂传灯周田田陈凯歌马广玉
Owner YANGZHOU UNIV
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