武汉市输入性恶性疟原虫裂殖子表面蛋白1、2基因分型研究  

Genotyping of merozoite surface protein 1 and 2 of imported Plasmodium falciparum in Wuhan City

作　　者：贾西帅 周水茂[1] 杨燕[1] 徐明星[1] 吴凯[1] 

机构地区：[1]武汉市疾病预防控制中心,武汉430015

出　　处：《中国寄生虫学与寄生虫病杂志》2017年第5期434-438,444,共6页Chinese Journal of Parasitology and Parasitic Diseases

摘　　要：目的了解武汉市输入性恶性疟原虫(Plasmodium falciparum)裂殖子表面蛋白1(merozoite surface protein 1,MSP1)和MSP2的等位基因型特征。方法于2010-2015年采集武汉市从非洲疟疾流行区回国人员中恶性疟现症患者血样,收集患者流行病学资料。提取患者血样中的恶性疟原虫DNA,采用巢式PCR分别扩增恶性疟原虫MSP1、MSP2基因型特异性片段,分析各等位基因型的构成比和不同感染来源地患者的基因型频数分布,分析重症患者不同基因型的相关性。不同基因型的重症患者比例差异采用χ~2检验。结果共采集输入性恶性疟患者血样175份,其中轻度症状患者血样137份,重症患者血样38份。巢式PCR结果显示,MSP1等位基因K1、RO33、MAD20型分别扩增出260~390、270和150 bp目的条带;MSP2等位基因3D7、FC27型分别扩增出200~330、330~500 bp目的条带。MSP1和MSP2基因均未检出的有9份。检出MSP1等位基因136份,检出率为77.7%,等位基因MAD20、K1、RO33、MAD20+K1、MAD20+RO33、K1+RO33、MAD20+K1+RO33型的构成比分别为5.1%(7/136)、37.5%(51/136)、20.6%(28/136)、5.9%(8/136)、4.4%(6/136)、16.9%(23/136)、9.5%(13/136)。检出MSP2等位基因143份,检出率为81.7%,FC27、3D7和FC27+3D7型的构成比为20.2%(29/143)、39.9%(57/143)、39.9%(57/143)。MAD20型和K1以南非最多,分别占10.5%(4/38)和34.2%(13/38);RO33型以东非最多,占2/7;FC27型东非和北非均未检出,南非占比最高,为18.4%(7/38);3D7型以东非最多,占4/7。MSP1基因MAD20、K1、RO33、MAD20+K1、MAD20+RO33、K1+RO33、MAD20+K1+RO33型的重症患者分别占1/7、19.6%(10/51)、32.1%(9/28)、1/8、4/6、13.0%(3/23)、46.2%(6/13),MAD20+RO33、MAD20+K1+RO33和RO33(P>0.05),与其他4种基因型间差异有统计学意义(P<0.05)。MSP2基因3D7、FC27、FC27+3D7型的重症患者比例分别为19.3%(11/57)、20.7%(6/29)、24.6%(14/57),各基因型间差异无统计学意义(P>0.05)。结论武汉市输入性恶性疟原虫MSP1存在MAD20、Kl和R033等3Objective To identify the genotypes of merozoite surface protein 1 （MSP1） and merozoite surface protein 2 （MSP2） of imported Plasmodium falciparum in Wuhan. Methods Blood samples were collected from re- turnees infected with P. falciparum from endemic areas of Africa, from 2010 to 2015 in Wuhan City, and epidemio- logical data were collected. The Plasmodium falciparum DNA was extracted from the blood samples. Nested PCR was used to amplify gene fragments of MSP1 and MSP2. The constituent ratios of the allelic families, the frequencies of each allelic family in patients from different countries, and the association of different allelic families with disease severity were analyzed. Analysis of proportions of severe cases with different genotypes were analyzed with A3 test. Results A total of 175 blood samples of patients with falciparum malaria were collected, which including 38 severe cases and 137 mild cases. Nested PCR for MSP1 allelic families K1, RO33 and MAD20 produced bands of 260- 390, 270 and 150 bp, and MSP2 allelic families 3D7 and FC27 produced bands of 200-330 and 330-500 bp. A total of 175 patients with falciparum malaria were involved in this study, of whom 9 patients showed negative results for both MSP1 and MSP2 amplification. The MSP1 allelic families were detected in 136 patients, with a detection rate of 77.7%. The constituent ratios of MAD20, K1, RO33, MAD20 ＋ K1, MAD20 ＋ RO33, K1 ＋ RO33, and MAD20 ＋ K1 ＋ RO33 were 5.1% （7/136）, 37.5% （51/136）, 20.6% （28/136）, 5.9% （8/136）, 4.4% （6/136）, 16.9% （23/136）, and 9.5% （13/136）, respectively. The MSP2 allelic families were detected in 143 patients, with a detection rate of 81.7%. The constituent ratios of FC27, 3D7, and FC27 ＋ 3D7 were 20.2% （29/143）, 39.9% （57/143）, and 39.9% （57/143）, re- spectively. The MAD20 allelotype was predominantly from South Africa （10.5%, 4/38）, the K1 was predominantly from South Africa （34.2%, 13/38）, and the RO33 was predominantly from East Africa （2/7

关 键 词：恶性疟原虫 裂殖子表面蛋白1 裂殖子表面蛋白2 基因分型 

分 类 号：R382.312[医药卫生—医学寄生虫学]