Primers and application of duplex PCR for early and rapid detection of Streptococcus agalactiae and Streptococcus iniae

Abstract

The invention discloses a double PCR primer for early rapid detection of Streptococcus agalactiae and Streptococcus iniae and its application. By using the primers of the invention to detect samples, one-time and simultaneous specific detection of Streptococcus agalactiae and Streptococcus iniae in the sample can be realized. The detection method established by the present invention can detect that the genome content is 9.84×10 ‑5 ng / μL of Streptococcus agalactiae and 9.30×10 ‑ 5 ng / μL Streptococcus iniae, and the concentration of the bacterial solution is 2.76×10 2 cfu / mL of Streptococcus agalactiae and 2.51×10 2 cfu / mL of Streptococcus iniae. This method is fast and accurate in the detection process of tilapia samples, has high specificity, sensitivity, repeatability and stability, and can realize early molecular early warning of tilapia streptococcosis, and has broad application prospects. It provides accurate and effective means for the rapid diagnosis and epidemiological investigation of the disease.

Description

technical field [0001] The invention belongs to the field of microorganism detection, in particular to a primer for early and rapid detection of Streptococcus agalactiae and Streptococcus iniae by double PCR and its application. Background technique [0002] Tilapia is one of the most widely farmed aquatic products in the world. According to incomplete statistics, in 2014, China's tilapia aquaculture production reached 1.6985 million tons, accounting for about 32% of the world's total aquaculture production. In recent years, tilapia streptococcosis has become increasingly serious, which has caused serious economic losses to the tilapia aquaculture industry. Studies have shown that the main pathogens of tilapia streptococcosis are Streptococcus agalactiae and Streptococcus iniae, and the most serious damage in recent years is Streptococcus agalactiae. The pathogenic bacteria can infect a variety of freshwater fish such as sea bream and tilapia, resulting in high mortality. ...

AI Technical Summary

Problems solved by technology

Both Streptococcus iniae and Streptococcus agalactiae can cause fish septicemia, meningitis and other diseases. It is difficult to accurately judge which kind of Streptococcus is caused by the external symptoms and the colony and shape of the two pathogenic bacteria. This will seriously affect the effective management of the disease

[0003] At present, the conventional diagnostic methods for Streptococcus tilapia are difficult to meet the requirements for rapid diagnosis of infected fish samples. At the same time, some physiological reactions may change with changes in the external environment, which can easily lead to misjudgment.

Ordinary PCR can only detect one gene at a time, which may cause missed or false detections

Fluorescent antibody technology and ELISA are expensive and require skilled technical operations; LAMP has false positives; quantitative PCR requires expensive instruments, professional operations, and high costs.

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