Sensitive and Rapid Detection of Traditional Chinese Herbs by Loop-Mediated Isothermal Amplification Method and Real-Time Fluorescence Quantitative PCR
Braz. arch. biol. technol
;
61: e18160292, 2018. tab, graf
Artículo
en Inglés
| LILACS
| ID: biblio-974117
ABSTRACT
ABSTRACT Adulterant herbal materials are threats to import and export trade and consumer safety. In this study, we established a simple and rapid examination system for the detection of Phellodendron chinense Schneid. Two detection methods, real-time fluorescence quantitative PCR (real-time PCR) and loop-mediated isothermal amplification (LAMP), were developed for traditional Chinese medicine detection, and their specificity and sensitivity were compared. The DNA of P. chinense was extracted and its special periods amplified with designed primers. Real-time PCR and LAMP experiments were conducted to test the specificity of primers in contrast to other similar species. The template concentration was diluted from 101 ng/µL to 10-5 ng/µL in order to contrast sensitivity between real-time PCR and LAMP. Real-time PCR and Lamp method has shown specificity because P. chinense was positive as opposed to other negative similar species. The Lamp method could detect a limited DNA concentration of 10-4ng/µL in 60 minutes with same sensitivity to real-time PCR. The results indicate that real-time PCR and LAMP are sensitive, accurate and specific in detection of P. chinense. However, LAMP is more convenient and cast less time. What's more, expensive equipments are not necessary for LAMP detector. For a better detection, we suggest an establishment of a real-time PCR and LAMP method for TCM market supervision which depends on DNA barcode sequences and LAMP.
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LILACS (Américas)
Asunto principal:
Phellodendron
/
Reacción en Cadena en Tiempo Real de la Polimerasa
/
Medicina Tradicional China
Tipo de estudio:
Estudio diagnóstico
Idioma:
Inglés
Revista:
Braz. arch. biol. technol
Asunto de la revista:
Biologia
Año:
2018
Tipo del documento:
Artículo
País de afiliación:
China
Institución/País de afiliación:
Liaoning Inspection and Quarantine Bureau/CN
/
Minzu University of China/CN
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