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目的筛选日本血吸虫3种重要保虫宿主来源虫体的特征性差异表达基因,寻找可以用于鉴别这3种宿主来源虫体的遗传标记。方法日本血吸虫尾蚴人工感染黄牛、水牛和山羊,收集感染后49 d成虫,分别抽提RNA,逆转录成cD NA,体外转录为cRNA并进行片段化;采用定制的血吸虫芯片分别进行杂交,筛选每种宿主来源虫体的特征性差异表达基因。采用实时定量PCR法对所筛选差异表达基因的表达水平做进一步测定,验证芯片数据的结果,并对这些特征性差异表达基因进行验证。结果通过芯片杂交,分析筛选获得实验黄牛组来源虫体特征性差异表达基因3个,水牛组来源虫体4个,山羊组来源虫体7个。其中黄牛组2个、水牛组3个、山羊组5个特征性差异表达基因在另一批次3种宿主来源虫体的重复试验中得到验证。结论芯片杂交筛选出10个3种宿主来源虫体的特征性差异表达基因,这些基因可能可以作为这3种不同宿主来源虫体的遗传标记。
Objective To screen differentially expressed genes of Schistosoma japonicum from three important hosts of Schistosoma japonicum and find genetic markers that can be used to identify the three host-derived parasites. Methods Adult cercariae of Schistosoma japonicum were artificially infected with buffaloes, buffaloes and goats at 49 days after infection. RNA was extracted and cD NA was reverse transcribed into cRNA and transcribed into cRNA in vitro. Fragmentation was performed using a custom-made schistosome chip. Species of differentially expressed genes of host parasites. Real-time quantitative PCR method was used to further determine the expression level of the differentially expressed genes selected to verify the results of the chip data and to validate the differentially expressed genes. Results By chip hybridization, 3 differentially expressed genes, 4 buffalo-derived parasites and 7 goat-derived parasites were obtained. Two of the cattle, three buffalo and five goat were differentially expressed in the replicates of the other three host-derived parasites. Conclusion Chromosome hybridization screened 10 differentially expressed genes of three kinds of host-derived worms, which may be used as genetic markers of these three kinds of host-derived parasites.