非洲输入性卵形疟病例的疟原虫k13基因螺旋浆区序列多样性分析  

Sequence diversity analysis of plasmodium k13 in helicoplasmic region of imported Plasmodium ovale cases from Africa

作　　者：金行一[1] 张家祺 朱素娟[1] 徐卫民[1] 霍亮亮[1] 陈华良[2] JIN Xing-yi;ZHANG Jia-qi;ZHU Su-juan;XU Wei-min;HUO Liang-liang;CHEN Hua-liang(Hangzhou Municipal Center for Disease Control and Prevention,Hangzhou,Zhejiang 310021,China;不详)

机构地区：[1]杭州市疾病预防控制中心,浙江杭州310021 [2]浙江省疾病预防控制中心,浙江杭州310051

出　　处：《中国卫生检验杂志》2022年第9期1030-1033,1037,共5页Chinese Journal of Health Laboratory Technology

基　　金：杭州市农业与社会发展科研主动设计项目(2019010-1A12);杭州市卫生计生科技计划项目(2018A79)。

摘　　要：目的为了解非洲输入性卵形疟原虫的二态性,对杭州市确诊的卵形疟病例感染疟原虫的k13基因多态性进行了分析。方法收集2014年2月—2021年6月杭州市诊断、报告的卵形疟现症病例血样22份,根据流行病学确认虫株感染地。提取疟原虫DNA,巢式PCR扩增、测序卵形疟原虫k13基因螺旋浆区206 aa~725 aa的编码区。测序序列与参比序列比对、整理获得cDNA序列,分析cDNA序列的单倍体类型和突变位点。结果卵形疟现症病例血样22份,90.9%(20/22)扩增到卵形疟原虫k13基因224 aa~725 aa编码区的DNA,测序、整理获得cDNA序列16条,存在3种单倍型,Curtisi单倍型、Wallikeri单倍型、突变型的占比分别为31.2%(5/16)、62.5%(10/16)和6.3%(1/16)。Curtisi型与Wallikeri型的序列之间存在c.711等38个碱基替换位点,突变型序列较之Wallikeri型,在c.1998位点出现T>A的同义突变。结论非洲输入杭州的卵形疟原虫以Wallikeri型为主,且突变型与Wallikeri型同源性更高。Objective In order to understand the dimorphism ofP.ovaleimported from Africa,polymorphisms of thek13 gene ofP.ovaleinfected patients in Hangzhou were analyzed.Methods Twenty-two blood samples were collected from both diagnosed and recorded cases in Hangzhou from February 2014 to June 2021,and original infection place was confirmed according to malaria epidemiology.For each sample the genomic DNA was extracted and the fragment encoding 206 aa-725 aa ofk13 gene propeller was amplified by nested PCR and sequenced.The cDNA sequence was obtained by aligning with the reference gene to identify the point mutations and haplotypes of cDNA sequence.Results The fragment of DNA encoding 224 aa-725 aa ofk13gene propeller was successfully amplified from 20 out of total 22 samples(90.9%).Eventually 16 cDNA sequences were obtained for analysis and 3 haplotypes were revealed.Among 3 haplotypes,the proportion of Curtisi haplotype,Wallikeri haplotype and mutant was 31.2%(5/16),62.5%(10/16)and 6.3%(1/16),respectively.There were 38 base substitution sites,such as c.711,between Curtisi and Wallikeri.Compared with Wallikeri,T>A synonym mutation was found in c.1998.Conclusion Wallikeri was the predominant genotype of importedPlasmodium ovaleisolates from Africa in Hangzhou.And the mutant haplotype showed high homogeneity to Wallikeri type.

关 键 词：非洲 输入性 卵形疟 疟原虫 k13基因螺旋浆区 多态性 

分 类 号：R531.3[医药卫生—内科学]