A triple fluorescent quantitative PCR detection method for detecting streptococcus agalactiae, streptococcus iniae and streptococcus gordonii

By employing a triple quantitative PCR method and utilizing the design of specific targets and primer probes, the problem of traditional PCR detection techniques being unable to efficiently detect Streptococcus agalactiae, Streptococcus dolphinii, and Lactococcus gasseri in aquaculture has been solved, enabling efficient and accurate identification of mixed infections and large-scale rapid screening.

CN122357752APending Publication Date: 2026-07-10CHINA AGRI UNIV SANYA RES INST
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
CHINA AGRI UNIV SANYA RES INST
Filing Date
2026-06-09
Publication Date
2026-07-10

AI Technical Summary

Technical Problem

Traditional PCR testing technology cannot efficiently, rapidly, and accurately detect Streptococcus agalactiae, Streptococcus dolphinus, and Lactococcus gasseri in aquaculture, especially in cases of mixed infection, and there is a risk of false positives, which cannot meet the needs of large-scale rapid screening.

Method used

This study employs a triple quantitative real-time PCR method, utilizing the neuB gene, simA gene, and the 16S-23S rDNA internal transcribed spacer (ITS) as detection targets. Specific primers and probes were designed, and combined with multiplex PCR technology, to simultaneously detect three pathogens in a single reaction system. PCR reaction conditions were optimized to reduce interference. Kits and detection methods are provided.

Benefits of technology

It achieves high specificity, low cost, and rapid detection of three pathogens, can identify mixed infections, improves detection efficiency and accuracy, reduces false positives, is suitable for large-scale rapid screening, and has important clinical guiding significance.

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Abstract

This invention discloses a triplet real-time PCR detection method for Streptococcus agalactiae, Streptococcus dolphinus, and Lactococcus gasseri. It utilizes three primer sets and probes: one for Streptococcus agalactiae, one for Streptococcus dolphinus, and one for Lactococcus gasseri. By optimizing the degradation temperature, primer dosage, and probe dosage, this invention effectively reduces the mutual interference of the three target pathogens, improving the accuracy of the detection results. This detection method has significant application value in fish farming. The reagent kit of this invention has low preparation cost, a simple preparation process, and is easily scalable for industrial production, thus possessing broad market prospects.
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