输入性卵形疟原虫细胞色素b、细胞色素c氧化酶Ⅰ和乳酸脱氢酶基因单核苷酸多态性分析  

Single nucleotide polymorphism analysis of cytochrome b,cytochrome c oxidase Ⅰ and lactate dehydrogenase genes of imported Plasmodium ovale

作　　者：张玲玲[1] 姚立农[1] 陈华良[1] 陆巧绎[1] 阮卫[1,2] 

机构地区：[1]浙江省疾病预防控制中心,杭州310051 [2]温州市卫生和计划生育委员会,温州325000

出　　处：《中国寄生虫学与寄生虫病杂志》2017年第5期429-433,共5页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：浙江省医药卫生科技项目(No.2014KYB324)~~

摘　　要：目的对卵形疟原虫(Plasmodium ovale)curtisi亚种和wallikeri亚种编码的细胞色素b(cytochrome b,Cytb),细胞色素c氧化酶Ⅰ(cytochrome c oxidaseⅠ,CoxⅠ)和乳酸脱氢酶(lactate dehydrogenase,LDH)基因的单核苷酸多态性进行分析。方法选择浙江省疟疾诊断参比实验室2011-2014年确诊的卵形疟原虫感染患者血样,提取卵形疟原虫DNA,巢式PCR扩增卵形疟原虫Cytb、CoxⅠ和LDH基因的部分片段,对扩增产物进行双向测序。测序序列经校对、拼接后,应用MEGA软件对测序序列和相应的氨基酸序列进行比对,分析单核苷酸多态性位点以及氨基酸突变位点。结果 19例卵形疟原虫感染者(其中curtisi亚种11例,wallikeri亚种8例)均为赴非洲返国人员。PCR扩增Cytb、CoxⅠ和LDH基因的产物长度分别为735、1 323和355 bp。curtisi和wallikeri两个亚种在Cytb、CoxⅠ和LDH基因分别发现10、13、13个单核苷酸二态性位点,且大多数的二态性位点属于同义突变。此外,两个亚种内部也存在核苷酸多态性,同时发现1例血样的CoxⅠ基因存在693个碱基的缺失。结论对卵形疟患者血样的Cytb、CoxⅠ和LDH基因进行了单核苷酸多态性分析,丰富了卵形疟原虫基因多态性信息。Objective To analyze the single nucleotide polymorphism （SNP） of cytochrome b （Cytb）, cytochrome c oxidase I （Cox I ） and lactate dehydrogenase （LDH） genes of Plasmodium ova/e curtisi and P. ova/e wa/likeri. Meth- ods The P. ova/e samples identified in Zhejiang Provincial Reference Laboratory for Malaria Diagnosis during 2011-2014 were used for nested PCR to amplify the partial fragments of Cytb, Cox I and LDH genes. The amplified products were sequenced in both directions. The nucleotide sequences and their corresponding amino acid sequences were aligned and analyzed using the Mega software to find out the SNP sites and amino acid mutation sites. Results The 19 samples of P. ova/e （11 curtisi and 8 wa/likeri） were all from persons returning from Africa. The lengths of PCR products for Cytb, Cox I and LDH were 735, 1 323 and 355 bp, respectively. There were 10, 13 and 13 single nucleotide dimorphism sites in Cytb, Cox ] and LDH, respectively, and most were synonymous mutation. Meanwhile, some nucleotide polymorphisms were found between the two subspecies. The Cox I gene was found to harbor a 693 bp deletion mutation in one sample. Conclusion We performed SNP analysis on Cytb, Cox I and LDH genes of P. ova/e, which will enrich the knowledge on gene polymorphism of P. ova/e.

关 键 词：卵形疟原虫 单核苷酸多态性 巢式PCR 

分 类 号：R382.31[医药卫生—医学寄生虫学]