MULTIPLEX POLYMERASE CHAIN-REACTION FOR DETECTION AND DIFFERENTIATION OF THE MICROBIAL INSECTICIDE BACILLUS-THURINGIENSIS
MULTIPLEX POLYMERASE CHAIN-REACTION FOR DETECTION AND DIFFERENTIATION OF THE MICROBIAL INSECTICIDE BACILLUS-THURINGIENSIS
复制标题
DOI：
10.1128/aem.59.2.523-527.1993
复制
发表时间：
1993-02-01
影响因子：
4.4
通讯作者：
LEVESQUE, RC
中科院分区：
文献类型：
--
作者：
BOURQUE, SN;VALERO, JR;LEVESQUE, RC
建立了一种快速鉴定苏云金芽孢杆菌的多重PCR方法。特异于编码三种主要类型B的基因内区域的高同源性引物。苏云金杆菌晶体蛋白用于产生B的每种菌株的PCR产物特征谱。苏云金kurstaki。根据PCR产物的电泳图谱对这些菌株进行区分。已知的B。苏云金通过PCR分析从昆虫尸体分离的kurstaki菌株以及未鉴定的菌株。在两步PCR中测定少量的粗样品裂解物，所述两步PCR含有能够区分分别产生CryIA（a）、CryIA（B）和CryIA（c）基因的1，500、858和653 bp产物的菌株的五种引物。该方法可用于B菌的快速检测。苏云金kurstaki在商业制剂和在该领域中的应用。
A rapid identification of Bacillus thuringiensis strains was established by using multiplex polymerase chain reaction (PCR). Primers of high homology specific to regions within genes encoding three major classes of B. thuringiensis crystal proteins were used to generate a PCR product profile characteristic of each strain of B. thuringiensis subsp. kurstaki. Differentiation among these strains was made on the basis of the electrophoretic pattern of the PCR products. Known B. thuringiensis subsp. kurstaki strains as well as unidentified strains isolated from insect cadavers were analyzed by PCR. Small amounts of crude sample lysates were assayed in a two-step PCR containing five primers capable of distinguishing between the strains giving products of 1,500, 858, and 653 bp for the CryIA(a), CryIA(b), and CryIA(c) genes, respectively. The method can be applied to rapidly detect the strains of B. thuringiensis subsp. kurstaki in commercial formulations and in the field.