Rapid authentication of Bupleurum species using an array of immobilized sequence-specific oligonucleotide probes.
Standardization and modernization of Chinese medicinal herbs are limited partially by misidentification of processed materials. Our goal was to develop an efficient method for verification of Chinese medicinal herbs, based on the variable sites of the rDNA internal transcribed spacer (ITS) region. We analyzed sequence differences in ITS of three Bupleurum species, B. kaoi Liu Chao et Chuang, B. falcatum L. and B. chinense DC., and developed a rapid detection method using a sequence-specific oligonucleotide probe (SSOP) array. The SSOP array, composed of poly-T tailed sequence-specific oligonucleotides, was hybridized to the digoxigenin (DIG)-labeled target ITS DNA of the Bupleurum species. The detected signals corresponded precisely to the specific sequences. This array provides a reliable and economical method for authenticating a large number of Chinese medicinal herbs. The short duration of the procedure (within 30 h) makes it an especially useful tool in verifying processed plant material.
Lin
Wen-Yu
WY
Institute of Bioinformatics and Structural Biology and Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan, Republic of China.
Chen
Li-Ru
LR
Lin
Tsai-Yun
TY
D001483:Base Sequence; D029972:Bupleurum / Q000145:classification* / Q000235:genetics*; D021901:DNA, Intergenic / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D013045:Species Specificity
D021901:DNA, Intergenic; D018744:DNA, Plant; D015345:Oligonucleotide Probes