Background & objectives: Malaria infects around 216 million people annually with estimated 445,000 deaths globally. Anopheles culicifacies is the vector of malaria in Sri Lanka, a complex of five morphologically identical sibling species of which precise identification using DNA–based methods is still under experimentation. This study was carried out in Sri Lanka to observe the utility of BCE-PCR assay based on mitochondrial Cytochrome Oxidase II (COII) developed in India, in sibling species B and E identification in Sri Lanka, to characterize nucleotide and corresponding amino acid sequences of COII region in major vector sibling species E in Sri Lanka and to analyze the spatial distribution pattern of sibling species E in Sri Lanka using microsatellite markers.
Methods: BCE-PCR was carried out for the samples to identify their sibling status. Sequencing of COII region was then carried out to investigate the genetic diversity of Sri Lankan sibling species E, sequences were aligned and compared; microsatellite genotyping was carried out and the spatial clustering pattern was analyzed.
Results: Identification of sibling species B and E using BCE-PCR was confusing due to the heterogeneity in the COII region of sibling species in Sri Lanka. Non-synonymous substitutions were detected in COII gene amongst sibling species E. Spatial distributed two clusters were detected in the studied population.
Interpretation & conclusion: Existence of genetic variants among sibling species is suggested in Sri Lanka. Further, the pattern of sibling species identification in BCE-PCR was reflected in the spatial clustering of sibling E in Sri Lanka.

中文翻译：

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线粒体-COII序列多态性反映斯里兰卡按蚊同胞种E的空间遗传聚类

背景和目标：疟疾每年感染约 2.16 亿人，全球估计有 445,000 人死亡。按蚊是斯里兰卡疟疾的载体，这是一种由五个形态相同的同胞物种组成的复合体，使用基于 DNA 的方法对其进行精确识别仍在试验中。本研究在斯里兰卡进行，以观察基于印度开发的线粒体细胞色素氧化酶 II (COII)的 BCE-PCR 测定在斯里兰卡的同胞物种 B 和 E 鉴定中的效用，以表征其核苷酸和相应的氨基酸序列。信息中心斯里兰卡主要媒介同胞种 E 的区域，并利用微卫星标记分析斯里兰卡同胞种 E 的空间分布格局。
方法：对样本进行 BCE-PCR，以确定其同胞状态。然后对COII区进行测序，考察斯里兰卡同胞种E的遗传多样性，进行序列比对和比较；进行了微卫星基因分型并分析了空间聚类模式。
结果：由于斯里兰卡同胞物种COII区域的异质性，使用 BCE-PCR 鉴定同胞物种 B 和 E 令人困惑。在COII中检测到非同义替换基因在兄弟物种 E. 空间分布的两个集群中检测到研究的种群。
解释和结论：在斯里兰卡建议同胞物种之间存在遗传变异。此外，BCE-PCR 中同胞物种鉴定的模式反映在斯里兰卡同胞 E 的空间聚类中。