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Molecular authentication of Gekko gecko using multienzyme isothermal rapid amplification technology
Jinjun Chai1, Yating Zhang1, Yuxin Xie1
1School of Medical Technology, Beihua University, Jilin, 132013, China.
Analytica Chimica Acta
|January 9, 2026
Summary
Two new methods, multienzyme isothermal rapid amplification combined with quantitative polymerase chain reaction (MIRA-qPCR) and MIRA with lateral flow dipstick (MIRA-LFD), rapidly authenticate Chinese medicinal ingredients like Gekko gecko. These simple, fast, and accurate techniques ensure the quality of traditional medicines.
Area of Science:
- Pharmacology and Traditional Chinese Medicine
- Molecular Biology and Genetics
- Biotechnology and Diagnostics
Background:
- Accurate authentication of Chinese medicinal materials is crucial for ensuring therapeutic efficacy and patient safety.
- Gekko gecko (G. gecko) is a valuable ingredient in Traditional Chinese Medicine, but is susceptible to adulteration and misidentification.
- Existing authentication methods can be time-consuming or require specialized equipment.
Purpose of the Study:
- To develop and validate two rapid and reliable methods for the authentication of Gekko gecko.
- To establish multienzyme isothermal rapid amplification combined with quantitative polymerase chain reaction (MIRA-qPCR) for G. gecko identification.
- To establish MIRA combined with lateral flow dipstick (MIRA-LFD) for rapid, on-site G. gecko authentication.
Main Methods:
- Design of specific primers and probes targeting the cytochrome oxidase I (COI) gene sequence of G. gecko.
- Optimization of reaction conditions for both MIRA-qPCR and MIRA-LFD systems.
- Evaluation of specificity, sensitivity, and speed of the developed MIRA-based methods.
Main Results:
- Both MIRA-qPCR and MIRA-LFD methods achieved amplification within 20 minutes at 39°C.
- MIRA-qPCR detected amplified fluorescent signals by the 8th cycle (4 minutes), while MIRA-LFD provided visual results in 5 minutes (optimal at 8 minutes).
- Both methods demonstrated high specificity, no cross-reactivity with common adulterants, and high sensitivity (10 pg/μL for MIRA-qPCR, 1 pg/μL for MIRA-LFD). Results from commercial samples were consistent.
Conclusions:
- MIRA-qPCR and MIRA-LFD are simple, fast, and accurate methods for G. gecko authentication.
- These methods show strong potential for routine application in quality control of Chinese medicinal ingredients.
- The developed techniques can be adapted for identifying other endangered or easily confused medicinal materials, enhancing reliability and efficacy.
