中藥鑒定是中藥質(zhì)量控制系統(tǒng)的首要環(huán)節(jié),中藥鑒定技術(shù)的不斷發(fā)展提高了藥材品質(zhì)評價水平.至今,中藥鑒定技術(shù)經(jīng)歷著從傳統(tǒng)生藥鑒定學(xué)的細(xì)胞和亞細(xì)胞水平向遺傳物質(zhì)DNA分子水平發(fā)展的歷程,運用DNA芯片技術(shù)進行中藥鑒定成為中藥鑒定發(fā)展的方向之一.介紹了篩選gDNA種特異性探針的抑制性差減微陣列"SSH-array"新技術(shù),該技術(shù)是將抑制性差減雜交與基于尼龍膜微陣列雜交相結(jié)合.以名貴中藥材石斛5個品種作為實驗材料,首先是用抑制性差減雜交獲得2個品種之間的gDNA差異片段,然后用這些gDNA差異片段與由所有研究品種的gDNA制備成的微陣列雜交篩選到某-個品種的種特異性gDNA片段.這些種特異性gDNA探針作為種特異性探針進行物種鑒定.在此基礎(chǔ)上建立中藥鑒定gDNA芯片的方法并對其應(yīng)用進行了展望探討.

The identification of traditional Chinese herbs(TCH)is very important in the quality control system of TCH.The development of theTCH identification techniques has improved the quality evaluation level of TCH.By so far.the TCH identification techniques have been developed from traditional pharmacognostic identification methods based on cell and sub-cell to genetic DNA(gDNA)level.It is a tend-ency to use DNA microarray to identify the species of TCH in recent years.In this review,a new method called suppression subtraction hybridization and DNA array(SSH-array)was introduced,which combined the SSH technique with microarray hybridization based on nylon film.Five plants fromDendrobium Sw.were used as experimental materials.Firstly the differential gDNA fragments between two species by SSH-array were obtained and then the diferential gDNA fragments with the microarrays made up of multiple whole genomes from several species were hybridized to screen the uniqued gDNA fragments for one species.The screened unique gDNA fragment can be used as species-specific probes for identifying the species.Based on the method,a procedure for species identification of TCH was established with gDNA microarray and its applied prospects were discussed.

國家自然科學(xué)基金資助項目(30371750);國家高技術(shù)研究計劃(2001AA2Z2012,2002AA2Z2041)