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首页> 外文期刊>Parasitology >A restriction site to differentiate Plasmodium and Haemoproteus infections in birds: on the inefficiency of general primers for detection of mixed infections.
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A restriction site to differentiate Plasmodium and Haemoproteus infections in birds: on the inefficiency of general primers for detection of mixed infections.

机译：区分鸟类疟原虫和血红蛋白感染的限制位点：检测混合感染的通用引物效率低下。

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摘要

Avian Plasmodium and Haemoproteus parasites are easily detected by DNA analyses of infected samples but only correctly assigned to each genus by sequencing and use of a phylogenetic approach. Here, we present a restriction site to differentiate between both parasite genera avoiding the use of those analyses. Alignments of 820 sequences currently listed in GenBank encoding a particular cytochrome B region of avian Plasmodium and Haemoproteus show a shared restriction site for both genera using the endonuclease Hpy CH4III. An additional restriction site is present in Plasmodium sequences that would initially allow differentiation of both genera by differential migration of digested products on gels. Overall 9 out of 326 sequences containing both potential restriction sites do not fit to the general rule. We used this differentiation of parasite genera based on Hpy CH4III restriction sites to evaluate the efficacy of 2 sets of general primers in detecting mixed infections. To do so, we used samples from hosts infected by parasites of both genera. The use of general primers was only able to detect 25% or less of the mixed infections. Therefore, parasite DNA amplification using general primers to determine the species composition of haemosporidian infections in individual hosts is not recommended. Specific primers for each species and study area should be designed until a new method can efficiently discriminate both parasites.
机译：通过感染样本的DNA分析可以轻松检测到禽疟原虫和变形杆菌的寄生虫，但是只有通过测序和系统进化方法才能正确地将其分配给每个属。在这里，我们提出了一个限制位点，以区分这两个寄生虫属，从而避免使用那些分析方法。使用核酸内切酶Hpy CH4III，当前在GenBank中列出的编码禽疟原虫和血红蛋白的特定细胞色素B区的820个序列的比对显示了两个属的共享限制位点。在疟原虫序列中存在另一个限制性位点，其最初将通过消化产物在凝胶上的差异迁移而允许两个属的分化。包含两个潜在限制位点的326个序列中总共有9个不符合一般规则。我们使用基于Hpy CH4III限制性位点的这种寄生虫属分化来评估2套通用引物在检测混合感染中的功效。为此，我们使用了被两个属的寄生虫感染的宿主的样本。使用通用引物只能检测到25％或更少的混合感染。因此，不建议使用通用引物进行寄生虫DNA扩增以确定单个宿主中血友病感染的物种组成。应针对每种物种和研究区域设计特定的引物，直到采用新方法可以有效区分两种寄生虫为止。

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