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使用任意引物PCR分析法检测恶性疟原虫的基因组多态性。

Detection of Genomic Polymorphism in Plasmodium falciparum using an arbitrarily primed PCR assay.

作者信息

Rojas M O, De-Castro J, Mariño G, Wasserman M

机构信息

Faculty of Sciences Universidad Nacional, Instituto Nacional de Salud, Bogotá, Colombia.

出版信息

J Eukaryot Microbiol. 1996 Jul-Aug;43(4):323-6. doi: 10.1111/j.1550-7408.1996.tb03995.x.

Abstract

Modifications of the arbitrarily primed polymerase chain reaction assay (i.e. a low annealing temperature and a very slow increase in the temperature during the elongation steps during the amplification cycles) allowed it to be used with the AT-rich Plasmodium falciparum DNA. The analysis of the products by polyacrylamide-urea gels, after silver staining, resulted in high resolution and sensitivity. Eighteen single and six combined pairs of arbitrary primers were tested. Two produced polymorphic patterns complex enough to differentiate between close Colombian isolates in a single assay. This method may be useful in studying the distribution and migration of strains in endemic areas, and for identifying intralaboratory cross-contamination of cultures.

摘要

对任意引物聚合酶链反应检测方法进行改进(即在扩增循环的延伸步骤中采用低退火温度和非常缓慢的温度升高),使其能够用于富含AT的恶性疟原虫DNA。通过聚丙烯酰胺-尿素凝胶对产物进行分析,银染后具有高分辨率和高灵敏度。测试了18种单引物和6种组合引物对。其中两种产生的多态性模式足够复杂,能够在一次检测中区分哥伦比亚的相近分离株。该方法可能有助于研究流行地区菌株的分布和迁移,以及识别实验室内部培养物的交叉污染。

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