Primer combination, kit and detection method for simultaneously detecting three echinococcosis pathogens

By designing primer combinations with high specificity and sensitivity and using multiplex PCR methods, the problems of false positives and false negatives in echinococcosis detection have been solved, enabling efficient and convenient simultaneous detection of three echinococcosis pathogens, improving detection efficiency and accuracy, and making it suitable for etiological identification in clinical and laboratory settings.

CN121046541APending Publication Date: 2025-12-02SICHUAN PROVINCIAL CENT FOR DISEASE CONTROL & PREVENTION +3
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Patent Information

Application Number
CN202511252843.5
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-09-03
Publication Date
2025-12-02

AI Technical Summary

Technical Problem

Existing methods for detecting echinococcosis suffer from false positives and false negatives, and primers are prone to cross-reaction during multiplex PCR, resulting in low detection efficiency and high costs, making it difficult to achieve efficient and convenient simultaneous detection of the three echinococcosis pathogens.

Method used

A primer combination with high specificity and sensitivity was designed for multiplex PCR detection of Echinococcus granulosus, Echinococcus multilocularis, and Echinococcus shiquensis. The primer combination includes Em-F1, Em-R1, Eg-F2, Eg-R2, Es-F3, and Es-R3. The kit contains Taq DNA polymerase, dNTPs, MgCl2, etc. The PCR reaction conditions are 94℃ pre-denaturation for 3 min, 94℃ denaturation for 30 s, 56℃ annealing for 40 s, and 72℃ extension for 40 s, for a total of 35 cycles.

Benefits of technology

It enables efficient, simple, and rapid detection of three echinococcosis pathogens in feces and tissues, with high specificity and sensitivity, significantly improving detection efficiency and accuracy, and reducing detection costs and time. It is suitable for etiological identification in clinical and laboratory settings.

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Abstract

The invention discloses a primer combination, a kit and a detection method for simultaneously detecting three echinococcosis pathogens, and belongs to the technical field of parasitic pathogen detection. The primer combination comprises a primer sequence as shown in SEQ ID NO. 1-6. The primer combination comprises a primer sequence as shown in SEQ ID NO. The primer pair combination designed by the invention does not interfere with each other when multiple PCR amplification is carried out in the same system under the same condition, is strong in specificity and high in sensitivity, and is designed for echinococcosis pathogens (echinococcus multilocularis, narrow-sense echinococcus granulosus and echinococcus stone). On the basis, a multiplex PCR method capable of identifying narrow-sense echinococcus granulosus, echinococcus multilocularis and echinococcus stone in faeces and tissues is established, the method is high in specificity and sensitivity, has the advantages of high flux, simplicity, convenience and rapidness, can remarkably improve the working efficiency, reduces the time and economic cost, and is suitable for popularization and application. The method has a wide application prospect in the field of echinococcosis molecular epidemiological investigation.
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