PCR-based identification of individuals of Schistosoma japonicum representing different subpopulations using a genetic marker in mitochondrial DNA
PCR-based identification of individuals of Schistosoma japonicum representing different subpopulations using a genetic marker in mitochondrial DNA
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DOI：
10.1016/s0020-7519(99)00040-5
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发表时间：
1999-07-01
影响因子：
4
通讯作者：
McManus, DP
中科院分区：
文献类型：
--
作者：
Sorensen, E;Bogh, HO;McManus, DP
测定了日本血吸虫3个实验室分离株的线粒体NADH脱氢酶I基因片段（NDI）。代表分离株的序列（最初从中华人民共和国的安徽省和浙江省以及从菲律宾获得）的比较显示分离株间的序列变异为0.2-0.6%，并且没有分离株内的变异是圆形的。该序列还表明，虽然浙江和菲律宾分离物的扩增产物含有核酸内切酶RsaI的识别位点，但安徽分离物中不存在这样的位点。这是通过用RsaI消化来自许多个体蠕虫的扩增产物来测试的。设计了一个感染实验，以验证该标记在研究该菌群体生物学中的价值。japonicum为最终宿主。为此，使用了两种中国分离株。三组小鼠（A-C）首先暴露于初次感染，然后在实验的第4周和第7周攻毒感染。在A组中，用安徽分离株进行第一次感染，用浙江分离株进行另外两次感染，从而提供特异性的可检测队列。B组和C组分别用安徽分离株进行第二次和第三次感染。所有小鼠在最后一次攻击感染后5周进行灌注，随后从灌注的蠕虫的DNA扩增NDI并用RsaI消化。消化结果显示，A、B感染群中存在安徽和浙江分离株的混合种群，而C感染群中仅存在浙江蠕虫。我们的结论是，在NDI中RsaI位点的存在/缺失为S.日本的(C)1999年澳大利亚寄生虫学会由爱思唯尔科技有限公司出版。保留所有权利。
A mitochondrial NADH dehydrogenase I gene fragment (NDI) was sequenced for three laboratory maintained isolates of Schistosoma japonicum. Comparison of sequences representing the isolates (originally obtained from the Anhui and Zhejiang provinces of the People's Republic of China, and from the Philippines) revealed inter-isolate sequence variations of 0.2-0.6% and no intra-isolate variation was round. the sequences also indicated that while the amplification products of the Zhejiang and Philippine isolates contained a recognition site for the endonuclease RsaI, there was no such site in the Anhui isolate. This was tested by digesting amplification products from a number of individual worms with RsaI. Then an infection experiment was designed to test the value of this genetic marker for studies of the population biology of S. japonicum in the final host. For this, the two Chinese isolates were used. Three groups of mice (A-C) were exposed firstly to a primary infection and then challenge-infected at weeks 4 and 7 of the experiment. In group A the first infection was done with the Anhui isolate, and the two others with the Zhejiang isolate, thereby providing a specific, detectable cohort. In groups B and C the Anhui isolate was used for the second and third infection. All mice were perfused 5 weeks after the last challenge infection, and the NDI was subsequently amplified from DNA of the perfused worms and digested with RsaI. The digestion revealed that while infection groups A and B contained mixed populations of the Anhui and Zhejiang isolates, only Zhejiang worms were present in group C. We concluded that the absence/presence of the RsaI site in the NDI provides a useful marker for the delineation of cohorts of S. japonicum. (C) 1999 Australian Society for Parasitology Inc. Published by Elsevier Science Ltd. All rights reserved.