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Primers, kits and methods for detecting Nocardia spp.

A technology of Nocardia amberjack and a kit is applied in the field of microbial detection, which can solve the problems of economic loss in aquaculture industry, long growth cycle of Nocardia and poor accuracy, etc., and achieves short detection time and application scope. Broad, easy-to-judge effect

Active Publication Date: 2022-06-28
GUANGDONG OCEAN UNIVERSITY +2
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Problems solved by technology

According to incomplete statistics, there are nearly 20 species of fish infected with the disease, including economic fish such as pomfret (Trachinotus ovatus), large yellow croaker (Larimichthys crocea), snakehead (Channamaculata), and largemouth bass (Micropterus salmoides). The farming industry has caused large economic losses
In the past, the detection of Nocardia spp. in yellowtails mostly used the traditional bacterial isolation and culture method. Due to the long growth cycle of Nocardia spp., it took too long and the accuracy was not good. There is an urgent need for a simple and easy-to-operate method in aquaculture production. Rapid and efficient detection technique for Nocardia spp. in yellowtail

Method used

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  • Primers, kits and methods for detecting Nocardia spp.
  • Primers, kits and methods for detecting Nocardia spp.
  • Primers, kits and methods for detecting Nocardia spp.

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Embodiment Construction

[0014] The following examples may enable those skilled in the art to more fully understand the present invention, but do not limit the present invention in any way.

[0015] At a constant temperature of 37°C, the recombinase is used to tightly bind the primer DNA to form a polymer of the enzyme and the primer. When the primer searches for a sequence that is completely complementary to it on the template DNA, the single-stranded DNA binding protein (single-stranded DNA) With the help of binding, SSB), the template DNA is melted, and under the action of DNA polymerase, a new DNA complementary chain is formed, and the reaction product also increases exponentially, usually within 0.5h. Amplified fragments detected by gel electrophoresis. Compared with detection methods such as PCR, LAMP, and qPCR, the method is simpler and requires less experimental equipment, which can meet the needs of grassroots units for rapid detection of Nocardia amberjack.

[0016] The present application ...

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Abstract

The invention discloses a primer, a kit and a method for detecting Nocardia spp. The primers for detecting Nocardia spp. include upstream primers and downstream primers, the nucleotide sequence of the upstream primers is as shown in SEQ.ID.N0.1, and the nucleotide sequence of the downstream primers is as shown in SEQ.ID.N0.2 Show. The invention solves the problems that the existing yellowtail nocardia detection technology has a long period, high detection cost, inapplicability to on-site rapid detection, and the like. The primer provided by the invention has good specificity, high sensitivity and wide application range.

Description

technical field [0001] The invention relates to the field of microorganism detection, and more particularly, to a primer, a kit and a method for detecting Nocardia amberjack. Background technique [0002] Nocardia amberjack is classified in the phylum Firmicutes, Actinobacteria, Actinobacterales, Nocardiaceae, Nocardia ). It is one of the main pathogenic bacteria in the aquaculture industry. After infection, a large number of nodules are formed on the surface and internal organs of the fish, and it can also cause bleeding and ulcers on the surface of the fish, which seriously affects the quality of the adult fish. The incidence of natural infection of Nocardia amberjack can reach 15-30%, and in severe cases, it can reach 50-60%, and the average mortality rate is about 20%. According to incomplete statistics, there are nearly 20 species of infected fish, including ovoid pomfret (Trachinotus ovatus), large yellow croaker (Larimichthys crocea), snakehead (Channamaculata), lar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/6844C12Q1/04C12N15/11
CPCC12Q1/689C12Q1/6844C12Q2531/119C12Q2565/125
Inventor 夏洪丽莫静怡夏立群鲁义善樊慧敏龙梦汪志文喻大鹏陈文捷
Owner GUANGDONG OCEAN UNIVERSITY
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