Da Silva D, Spada R G, Sobral-Hamaguchi S S, Abdel-Hamid Z, Zuim N R, Zanotti-Magalhães E M, Magalhães L A, Ribeiro-Paes J T
Department of Biological Sciences, UNESP, Av. Dom Antonio, 2100, Assis, SP, Brazil, 19806-900.
Parasite. 2004 Mar;11(1):43-9. doi: 10.1051/parasite/200411143.
DNA analysis by molecular techniques has significantly expanded the perspectives of the study and understanding of genetic variability in molluscs that are vectors of schistosomiasis. In the present study, the genetic variability of susceptible and resistant B. lenagophila strains to S. mansoni infection was investigated using amplification of their genomic DNA by RAPD-PCR. The products were analyzed by PAGE and stained with silver. The results showed polymorphism between tested strains with four different primers. We found two bands of 1,900 and 3,420 bp that were characteristic of the susceptible strains with primer 2. The primers 9 and 10 identified a single polymorphic band that was also characteristic of (3,136 and 5,041 bp, respectively) susceptible snails. Two polymorphic bands were detected by primer 15: one with 1,800 bp was characteristic of the resistant strain and the other with approximately equal to 1,700 bp in the susceptible one. These results provide additional evidence showing that the RAPD-PCR technique is adequate for the study of polymorphisms in intermediate hosts snails of S. mansoni. The obtained results are expected to expand the knowledge about the genetic variability of the snails and to permit the future identification of genomic sequences specifically related to the resistance/susceptibility of Biomphalaria to the larval forms of S. mansoni.
通过分子技术进行的DNA分析显著拓宽了对作为血吸虫病传播媒介的软体动物遗传变异性的研究和理解视角。在本研究中,利用RAPD-PCR扩增易感和抗性嗜卷螺菌株的基因组DNA,研究了它们对曼氏血吸虫感染的遗传变异性。产物通过PAGE分析并用银染色。结果显示,使用四种不同引物时,受试菌株之间存在多态性。我们发现引物2扩增出的1900和3420 bp两条带是易感菌株的特征带。引物9和10分别鉴定出一条单一的多态性条带(分别为3136和5041 bp),这也是易感螺的特征带。引物15检测到两条多态性条带:一条1800 bp的带是抗性菌株的特征带,另一条在易感菌株中约为1700 bp。这些结果提供了额外的证据,表明RAPD-PCR技术适用于研究曼氏血吸虫中间宿主螺的多态性。预期所获得的结果将扩展有关螺遗传变异性的知识,并有助于未来鉴定与嗜卷螺对曼氏血吸虫幼虫形式的抗性/易感性具体相关的基因组序列。