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 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.

Comparison Copper Absorption Capacity with Several Bacteria Stains from the Surface of Macroalga Gracilaria lemaneaformis(Rhodophyta) / 微生物学通报

The main pollution of coastal sea is heavy metal and eutrophication,which resulted in degeneration of its eco-function.Pollution of heavy metal is accompanied with eutrophication in sea water.This article studied on copper absorption capacity of several bacteria strains which were isolated from the surface of macroalga Gracilaria lemaneaformis that was cultivated in copper polluted sea.We got 6 strains of epiphytic bacteria from the seaweed,Ochrobactrum anthropi,Aeromonas salmonicida,Micrococcus lylae,Corynebacterium ulcerans,Pseudoalteromonas spp,Vibrio fluvialis.Attained from the experiment of copper biosorption,A.salmonicida had a maximal biosorption capacity among them,and Vibrio minimal.Based on this trail,A.salmonicida,C.ulcerans and Vibrio were selected for next trails included optimal biosorption time,optimal pH value and effect of pre-disposal by several chemical reagents on biosorption capacity.The results showed:optimal biosorption time was between 40 minutes and 60 minutes,and optical pH value 4～5.Pre-disposal by HCl leaded to lost the absorption capacity of A.salmonicida,but was opposite by NaOH and ethanol.For C.ulcerans and Vibrio,the copper absorption capacities were increased by them.