恶性疟原虫组氨酸富集蛋白Ⅱ、Ⅲ的序列多态性分析  

Polymorphism Analysis of Plasmodium falciparum Histidine-rich Protein Ⅱ and Ⅲ

作　　者：杨英超[1] 张瑾[1] 王国柱[1] 薄淑英[1] 张影[1] 辛晓芳[1] 

机构地区：[1]中国食品药品检定研究院,卫生部生物技术产品检定方法及其标准化重点实验室,北京100050

出　　处：《中国寄生虫学与寄生虫病杂志》2014年第5期373-376,共4页Chinese Journal of Parasitology and Parasitic Diseases

摘　　要：目的分析恶性疟原虫富组氨酸蛋白Ⅱ/Ⅲ（Pf HRPⅡ/Ⅲ）的序列多态性。方法采集云南疟疾流行区恶性疟患者血样20份,血涂片后镜检,并进行疟疾快速诊断试剂检测（RDTs）,同时提取各血样中恶性疟原虫基因组DNA,进行PCR扩增Pfhrp2和Pfhrp3核酸片段并测序。使用Bioedit软件对Pfhrp2和Pfhrp3基因序列进行比对,使用TRANSEQ软件推导出其编码的氨基酸序列。结果 20份血样经镜检和RDTs检测均为恶性疟原虫阳性。PCR扩增结果显示,各血样的Pfhrp2核酸片段约389～986 bp,Pfhrp3核酸片段大小约329～640 bp,应用测序获得的核酸序列推导出相应的氨基酸序列,Pf HRPⅡ氨基酸残基序列以1型（AHHAHHVAD）起始并最终以12型（AHHAAAHHEAATH）作为结尾,Pf HRPⅡ特征性氨基酸残基重复序列数量相对较多的分别有7型（AHHAAD）、2型（AHHAHHAAD）和6型（AHHATD）;Pf HRPⅢ含有重复较多的型别是16型（AHHAAN）和17型（AHHDG）;Pf HRPⅡ和Pf HRPⅢ均未发现11型（AHN）。结论 20份恶性疟患者血样中Pfhrp2和Pfhrp3存在诸多的核酸变异,Pf HRPⅡ和Pf HRPⅢ共享一些特征性重复序列。Objective To analyze the polymorphism of Plasmodium falciparum histidine-rich protein （PfHRP） Ⅱ and Ⅲ. Methods Genomie DNA was isolated from blood samples of 20 patients infected with Plasmodium falciparum in Yunnan Province. Blood samples were tested by microscopy and RDTs. The Pfhrp2 and Pfhrp3 gene fragments were amplified by PCR and sequenced. The sequencing results were analyzed and compared using the bioinformatics software. Results 20 patients infected with Plasmodium falciparum tested by microscopy and RDTs. PCR showed that the Pfhrp2 gene was with 389--986 bp, and Pfhrp3 gene with 329～640 bp. All PfHRP Ⅱ sequences started with type 1 repeat （AHHAHHVAD） and ended with the type 12 repeat （AHHAAAHHEAATH）. The number of type 7 （AHHAAD）, type 2 （AHHAHHAAD） and type 6 （AHHATD） within PfHRPⅡ was more than the other types of repeats, as well as type 16 （AHHAAN） and type 17 （AHHDG） for Pfl-IRPⅢ. Type 11 repeat （AHN） was not found from the PfHRPⅡ and PfHRPⅢ sequences. Conclusion There is an extensive diversity in Pfhrp2 and Pfhrp3 fragments in the individuals infected with P.falciparum in Yunnan. Some types of repeats are shared by PfHRPⅡ and PfHRPⅢ.

关 键 词：恶性疟原虫 富组氨酸蛋白Ⅱ 富组氨酸蛋白Ⅲ 分型 

分 类 号：R382.312[医药卫生—医学寄生虫学]