McReynolds L A, Poole C, Hong Y, Williams S A, Partono F, Bradley J
Molecular Parasitology Group, New England Biolabs, Beverly, MA.
Southeast Asian J Trop Med Public Health. 1993;24 Suppl 2:55-63.
Monitoring of filarial parasites in the host and vector has traditionally depended on morphological identification. Recently, species-specific DNA probes have been developed for Brugia malayi, Brugia pahangi and Wuchereria bancrofti. Repeated DNA sequences are useful in developing DNA probes because they evolve more rapidly then coding sequences and their high copy number increases the sensitivity of detection. The Hhal repeated DNA family represents 12% of the total B. malayi DNA. This DNA family is present in species of Brugia (B. malayi, B. timori and B. pahangi) but not W. bancrofti. Sequence analysis of the repeated DNA in B. malayi and B. pahangi has allowed construction of two species-specific DNA probes. These probes were used in a double blind field study in Indonesia. Microfilariae (mf) from infected cats and humans were identified by classical morphological methods and DNA probes. Agreement was found in 98.6% of the 642 samples tested by the two different techniques. Besides mf identification DNA probes can be used to determine the species of infective larvae (L3s) in infected mosquitos. This is useful because the L3s have similar morphology. DNA probes for the identification of W. bancrofti have recently been developed and are in the initial stages of testing in China (Piessens, personal communication) and Egypt (Williams, personal communication). An alternative approach for identification of infected individuals is to detect specific parasite antigens in circulation. A WHO initiative to use either an antigen or antibody assay to replace night blood is presently underway. This approach, if successful would not require the presence of microfilariae, but could detect occult infections.
传统上,对宿主和媒介中丝虫寄生虫的监测依赖于形态学鉴定。最近,已开发出针对马来布鲁线虫、帕氏布鲁线虫和班氏吴策线虫的种特异性DNA探针。重复DNA序列在开发DNA探针时很有用,因为它们的进化速度比编码序列更快,且其高拷贝数增加了检测的灵敏度。Hhal重复DNA家族占马来布鲁线虫总DNA的12%。该DNA家族存在于布鲁线虫属物种(马来布鲁线虫、帝汶布鲁线虫和帕氏布鲁线虫)中,但不存在于班氏吴策线虫中。对马来布鲁线虫和帕氏布鲁线虫中重复DNA的序列分析使得能够构建两种种特异性DNA探针。这些探针在印度尼西亚的一项双盲现场研究中得到应用。通过经典形态学方法和DNA探针鉴定来自受感染猫和人类的微丝蚴。在两种不同技术检测的642个样本中,98.6%的结果一致。除了鉴定微丝蚴,DNA探针还可用于确定受感染蚊子中感染性幼虫(L3)的种类。这很有用,因为L3的形态相似。最近已开发出用于鉴定班氏吴策线虫的DNA探针,目前正在中国(皮森森,个人交流)和埃及(威廉姆斯,个人交流)处于测试的初始阶段。鉴定受感染个体的另一种方法是检测循环中的特异性寄生虫抗原。世界卫生组织目前正在开展一项倡议,使用抗原或抗体检测来取代夜间采血。如果这种方法成功,将不需要存在微丝蚴,但可以检测隐匿性感染。