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用随机扩增多态性DNA指纹技术鉴定和分类淋病奈瑟菌

Identification and classification of Neisseria gonorrhoeae by RAPD fingerprinting.

作者信息

Zhu X, Kong F, Zhang G, Chen S

机构信息

Department of Dermatology, First Hospital, Beijing Medical University.

出版信息

Chin Med J (Engl). 1995 Apr;108(4):269-72.

PMID:7789214
Abstract

Using the RAPD method, we identified and classified Neisseria gonorrhoeae on the basis of the DNA level. Except OPG20 the other 19 primers of OPG arbitrary primer kit showed informative arrays of amplified prominent segments. Most of the segments were common to all three Neisseria gonorrhoeae groups; some segments were amplified from two groups or only one group so that the fingerprint maps of different Neisseria gonorrhoeae groups were distinctive. Based on these findings, i.e. on the genomic level, we can classify the three Neisseria gonorrhoeae groups. Using OPG10 and OPG11 to amplify the five Neisseria species, we found marked diversity among them, which made it easy for us to identify Neisseria gonorrhoeae from four other Neisseria species. The use of the RAPD method needs some special reaction conditions, e.g. annealing temperature that should not be too low or too high and should be consistently stable. The quality of Taq polymerase is also very important. The reproducibility of the results under these conditions was very good.

摘要

我们运用随机扩增多态性DNA(RAPD)方法,在DNA水平上对淋病奈瑟菌进行鉴定和分类。除OPG20外,OPG随机引物试剂盒的其他19种引物均显示出信息丰富的扩增显著片段阵列。大多数片段在所有三个淋病奈瑟菌群中都很常见;一些片段仅从两个菌群或仅一个菌群中扩增出来,因此不同淋病奈瑟菌群的指纹图谱各不相同。基于这些发现,即在基因组水平上,我们可以对这三个淋病奈瑟菌群进行分类。使用OPG10和OPG11扩增五种奈瑟菌,我们发现它们之间存在显著差异,这使得我们很容易从其他四种奈瑟菌中鉴定出淋病奈瑟菌。RAPD方法的使用需要一些特殊的反应条件,例如退火温度不应过低或过高,且应始终保持稳定。Taq聚合酶的质量也非常重要。在这些条件下结果的可重复性非常好。

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