Sim B K, Piessens W F, Wirth D F
Mol Biochem Parasitol. 1986 May;19(2):117-23. doi: 10.1016/0166-6851(86)90116-7.
This report describes a specific and sensitive DNA probe for the identification of Brugia malayi. A genomic DNA library produced from subperiodic B. malayi microfilariae was screened to detect clones containing DNA sequences which are highly repeated within the parasite genome. Several clones were further analyzed to identify those which hybridize specifically with B. malayi DNA but not with DNA from B. pahangi and Dirofilaria immitis. From these, clone pBm15 was selected because it hybridized with high sensitivity to B. malayi DNA as detected by autoradiography. Clone pBm15 was sensitive enough to detect two infective larvae or five microfilariae or 300 pg of purified B. malayi microfilarial DNA. This study forms the basis for the development of a specific and sensitive DNA probe for the identification of B. malayi in field specimens.
本报告描述了一种用于鉴定马来布鲁线虫的特异性灵敏DNA探针。对由亚周期型马来布鲁线虫微丝蚴构建的基因组DNA文库进行筛选,以检测含有在寄生虫基因组中高度重复的DNA序列的克隆。对几个克隆进行进一步分析,以鉴定那些与马来布鲁线虫DNA特异性杂交但不与彭亨布鲁线虫和犬恶丝虫DNA杂交的克隆。从中选择了克隆pBm15,因为通过放射自显影检测,它与马来布鲁线虫DNA具有高灵敏度的杂交。克隆pBm15灵敏到足以检测两条感染性幼虫或五条微丝蚴或300 pg纯化的马来布鲁线虫微丝蚴DNA。本研究为开发用于鉴定野外标本中马来布鲁线虫的特异性灵敏DNA探针奠定了基础。