Morgan U M, Constantine C C, O'Donoghue P, Meloni B P, O'Brien P A, Thompson R C
World Health Organization Collaborating Centre for the Molecular Epidemiology of Parasitic Infections, School of Veterinary Studies, Murdoch University, Australia.
Am J Trop Med Hyg. 1995 Jun;52(6):559-64. doi: 10.4269/ajtmh.1995.52.559.
Genetic variation in 25 Cryptosporidium isolates was analyzed using the random amplified polymorphic DNA (RAPD) technique. Simple reproducible polymorphisms were generated (using five primers) from Cryptosporidium DNA that was free of contaminating bacterial DNA. The results generated by four of the five primers were statistically correlated (P < 0.001). The combined data from three primers were used to construct a phenogram using Jaccard's distance. Four groupings could be distinguished. Two C. serpentis isolates from snakes formed a distinct group of their own, whereas C. parvum isolates were divided into two main groups: one containing most human isolates and the other containing mostly domestic animals plus two remaining human isolates. Due to the sensitivity of the RAPD technique, isolates can now be analyzed genetically, directly from fecal samples without further biological amplification. This represents a significant advance on current techniques.
使用随机扩增多态性DNA(RAPD)技术分析了25株隐孢子虫分离株的遗传变异。从不含污染细菌DNA的隐孢子虫DNA中(使用五种引物)产生了简单可重复的多态性。五种引物中的四种产生的结果具有统计学相关性(P < 0.001)。使用来自三种引物的合并数据,利用杰卡德距离构建了一个系统树图。可以区分出四个分组。来自蛇的两株蛇隐孢子虫分离株形成了一个独特的组,而微小隐孢子虫分离株则分为两个主要组:一组包含大多数人类分离株,另一组主要包含家畜以及另外两株人类分离株。由于RAPD技术的敏感性,现在可以直接从粪便样本中对分离株进行基因分析,无需进一步的生物扩增。这代表了当前技术的重大进步。