Sensitive and Rapid Detection of Traditional Chinese Herbs by Loop-Mediated Isothermal Amplification Method and Real-Time Fluorescence Quantitative PCR
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...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Instituto de Tecnologia do Paraná (Tecpar)
2018-11-01
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| Series: | Brazilian Archives of Biology and Technology |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132018000100314&lng=en&tlng=en |
| _version_ | 1819096030572969984 |
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| author | Min Min Ma Dan Jiang Shuang Ya Li Yuan Wu Ju Meng Jiang Yao Huang |
| author_facet | Min Min Ma Dan Jiang Shuang Ya Li Yuan Wu Ju Meng Jiang Yao Huang |
| author_sort | Min Min Ma |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-21T23:52:43Z |
| format | Article |
| id | doaj.art-e5e2c801ea014145bf214ed4de1da1fe |
| institution | Directory Open Access Journal |
| issn | 1678-4324 |
| language | English |
| last_indexed | 2024-12-21T23:52:43Z |
| publishDate | 2018-11-01 |
| publisher | Instituto de Tecnologia do Paraná (Tecpar) |
| record_format | Article |
| series | Brazilian Archives of Biology and Technology |
| spelling | doaj.art-e5e2c801ea014145bf214ed4de1da1fe2022-12-21T18:45:53ZengInstituto de Tecnologia do Paraná (Tecpar)Brazilian Archives of Biology and Technology1678-43242018-11-0161010.1590/1678-4324-2018160292S1516-89132018000100314Sensitive and Rapid Detection of Traditional Chinese Herbs by Loop-Mediated Isothermal Amplification Method and Real-Time Fluorescence Quantitative PCRMin Min MaDan JiangShuang Ya LiYuan WuJu MengJiang Yao HuangABSTRACT 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.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132018000100314&lng=en&tlng=enTraditional Chinese medicineReal-time PCRPhellodendron chinense SchneidLAMPSpecificitySensitivity |
| spellingShingle | Min Min Ma Dan Jiang Shuang Ya Li Yuan Wu Ju Meng Jiang Yao Huang Sensitive and Rapid Detection of Traditional Chinese Herbs by Loop-Mediated Isothermal Amplification Method and Real-Time Fluorescence Quantitative PCR Brazilian Archives of Biology and Technology Traditional Chinese medicine Real-time PCR Phellodendron chinense Schneid LAMP Specificity Sensitivity |
| title | Sensitive and Rapid Detection of Traditional Chinese Herbs by Loop-Mediated Isothermal Amplification Method and Real-Time Fluorescence Quantitative PCR |
| title_full | Sensitive and Rapid Detection of Traditional Chinese Herbs by Loop-Mediated Isothermal Amplification Method and Real-Time Fluorescence Quantitative PCR |
| title_fullStr | Sensitive and Rapid Detection of Traditional Chinese Herbs by Loop-Mediated Isothermal Amplification Method and Real-Time Fluorescence Quantitative PCR |
| title_full_unstemmed | Sensitive and Rapid Detection of Traditional Chinese Herbs by Loop-Mediated Isothermal Amplification Method and Real-Time Fluorescence Quantitative PCR |
| title_short | Sensitive and Rapid Detection of Traditional Chinese Herbs by Loop-Mediated Isothermal Amplification Method and Real-Time Fluorescence Quantitative PCR |
| title_sort | sensitive and rapid detection of traditional chinese herbs by loop mediated isothermal amplification method and real time fluorescence quantitative pcr |
| topic | Traditional Chinese medicine Real-time PCR Phellodendron chinense Schneid LAMP Specificity Sensitivity |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132018000100314&lng=en&tlng=en |
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