Rapid identification of yeast species using three primers in a polymerase chain reaction.
Rapid identification of yeast species using three primers in a polymerase chain reaction.
复制标题
在聚合酶链式反应中使用三种引物快速鉴定酵母种类。
DOI：
--
复制
发表时间：
1993
期刊：
影响因子：
--
通讯作者：
J. Fell
中科院分区：
文献类型：
--
作者：
J. Fell
传统的酵母菌鉴定方法依赖于各种形态和生理测试，通常需要几天到几周的时间才能完成。我们已经能够通过在聚合酶链式反应中使用大亚基rDNA区域的多片段特异性寡核苷酸启动来将时间减少到不到一天。“热启动”反应采用2个通用的外部定界引物和1个内部的种属特异性引物。共检测了5条特异引物：一条针对红酵母生物相似类群的引物，一条属于胞菌属(Cystofilasidium)的引物，以及3条种特异的引物(白孢子菌、隐球菌属和粘红酵母)。在没有特定目标DNA的情况下，通用的rDNA片段被扩增；在有目标DNA存在的情况下，特定的引物区被扩增。当使用24个核苷酸特异的引物时，该技术的准确度在两个碱基位置的差异内。这项技术应该也适用于其他海洋真核生物。
Classic methods for identification of yeasts rely on a variety of morphological and physiological tests that often take days to weeks to complete. We have been able to reduce the time to less than one day through the use of multiple segment-specific oligonucleotide priming of a region of the large subunit rDNA in a polymerase chain reaction. The "hot start" reaction was used with two universal external delimiting primers and one internal species-specific primer. Five specific primers were tested: a primer for a biologically similar group of Rhodotorula species, a generic (Cystofilobasidium) primer, and 3 species-specific primers (Leucosporidium scottii, Cryptococcus muscorum, and Rhodotorula mucilaginosa). In the absence of specific target DNA, the universal rDNA segment is amplified; in the presence of target DNA, the specific primer region is amplified. The technique is accurate within two base position differences when a 24 nucleotide-specific primer is used. The technique should be applicable to other marine eukaryotes.