Polymorphism analysis of the block 5 region in merozoite surface protein-1 gene of imported and local Plasmodium vivax in Yunnan Province

Abstract

Abstract:

Objective To analyze the sequence of Plasmodium vivax merozoite surface protein-1 (PvMSP-1) and allele polymorphism in imported and local vivax malaria parasites in Yunnan Province. Methods Blood samples on filter paper were collected from imported and local vivax malaria cases in Yunnan Province during August 2012 and September 2015 and information of epidemiological history was recorded. Plasmodium DNA was extracted by a DNA extraction kit, and the block 5 region in PvMSP-1 gene was amplified by PCR. The PCR products were sequenced and blasted with reference sequences M75674, AAN86237, M60807, ABJ53045, AAN86238 and BAA18977. The sequence polymorphism in block 5 region of PvMSP-1 was analyzed with MEGA 5.04 and Arlequin3.5.1 softwares. The conserved sites, genetic distances among sequences and Shannon-wiener index among alleles were calculated. The clustering tree was drawn according to the genetic distances between the amino-acid sequences. Results A total of 847 blood samples were collected from the malaria cases, comprising of 61 samples from local cases, 66 from imported cases from Africa, and 720 from Myanmar. The block 5 region in PvMSP-1 was successfully amplified in 278 samples, and sequencing was successfully made in 206 of them. The peptide coded by the block 5 region had a length of 193 to 222 aa. The amino acid sequence alignment showed that in 206 samples the proportion of genotypes of Sal-1, Belem and Recombine was 59.2% (122/206), 23.3% （48/206） and 17.5% （36/206）, respectively. The proportion of Sal-1 genotype in imported cases from Myanmar and Africa and in local cases was 58.8% (104/177), 73.3% (11/15) and 50% (7/14), respectively. The genotypes Sal-1, Belem and Recombine had 51, 9 and 6 different alleles. The 66 alleles had a Shannon Wiener index (H’) of 0.955 and an expected heterozygosis (He) of 0.567. The 206 DNA sequences had a 665-bp homologous locus, comprising of 75 conserved sites (11.3%, 75/665) and 590 variable sites (88.7%, 590/665). The genetic distances between sequences were all less than 0.4. The clustering analysis showed that the 206 sequences were clustered into two categories with three branches. The homology of Recombine with Belem genotype was 91%-92%, higher than with Sal-1 genotype (82%-83%). Conclusion The block 5 region in PvMSP-1 gene from local and imported Plasmodium vivax in Yunnan Province has varied forms of alleles, and the Sal-1 genotype is predominant among the three genotypes.

Key words: Plasmodium vivax, Merozoite surface protein-1, Polymorphism

CLC Number:

R382.311

Ying DONG, Ai-ming SUN, Meng-ni CHEN, Yan-chun XU, Xiang-hua MAO, Yan DENG. Polymorphism analysis of the block 5 region in merozoite surface protein-1 gene of imported and local Plasmodium vivax in Yunnan Province[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2017, 35(1): 1-7.

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References 38

[1]	World Health Organization.World malaria report 2014[R]. Geneva: WHO, 2014: 3.
[2]	Carlton JM, Adams JH, Silva JC, et al. Comparative genomics of the neglected human malaria parasite Plasmodium vivax[J]. Nature, 2008, 455(7214): 757-763.
[3]	周水森, 王漪, 汤林华. 2005年全国疟疾形势[J]. 中国寄生虫学与寄生虫病杂志, 2006, 24(6): 401-403.
[4]	周水森, 王漪, 汤林华. 2006年全国疟疾形势[J]. 中国寄生虫学与寄生虫病杂志, 2007, 25(6): 439-441.
[5]	周水森, 王漪, 房文, 等. 2007年全国疟疾形势[J]. 中国寄生虫学与寄生虫病杂志, 2008, 26(6): 401-403.
[6]	周水森, 王漪, 房文, 等. 2008年全国疟疾形势[J]. 中国寄生虫学与寄生虫病杂志, 2009, 27(6): 455-457.
[7]	周水森, 王漪, 夏志贵. 2009年全国疟疾疫情分析[J]. 中国寄生虫学与寄生虫病杂志, 2011, 29(1): 1-3.
[8]	周水森, 王漪, 李雨. 2010年全国疟疾疫情分析[J]. 中国寄生虫学与寄生虫病杂志, 2011, 29(6): 401-403.
[9]	夏志贵, 杨曼尼, 周水森. 2011年全国疟疾疫情分析[J]. 中国寄生虫学与寄生虫病杂志, 2012, 30(6): 419-422.
[10]	夏志贵, 丰俊, 周水森. 2012年全国疟疾疫情分析[J]. 中国寄生虫学与寄生虫病杂志, 2013, 31(6): 413-418.
[11]	张丽, 丰俊, 夏志贵. 2013年全国疟疾疫情分析[J]. 中国寄生虫学与寄生虫病杂志, 2014, 32(6): 407-413.
[12]	张丽, 周水森, 丰俊, 等. 2014年全国疟疾疫情分析[J]. 中国寄生虫学与寄生虫病杂志, 2015, 33(5): 319-326.
[13]	Alecrim M das G, Alecrim W, Macêdo V. Plasmodium vivax resistance to chloroquine(R2) and mefloquine(R3) in Brazilian Amazon region[J]. Rev Soc Bras Med Trop, 1999, 32(1): 67-68.
[14]	Chehuan YF, Costa MR, Costa JS, et al. In vitro chloroquine resistance for Plasmodium vivax isolates from the Western Brazilian Amazon[J]. Malar J, 2013, 12: 226.
[15]	Leslie T, Mayan MI, Hasan MA, et al. Sulfadoxine-pyrimethamine, chlorproguanil-dapsone, or chloroquine for the treatment of Plasmodium vivax malaria in Afghanistan and Pakistan: a randomized controlled trial[J]. JAMA, 2007, 297(20): 2201-2209.
[16]	Cui L, Mascorro CN, Fan Q, et al. Genetic diversity and multiple infections of Plasmodium vivax malaria in Western Thailand[J]. Am J Trop Med Hyg, 2003, 68(5): 613-619.
[17]	Imwong M, Pukrittayakamee S, Grüner AC, et al. Practical PCR genotyping protocols for Plasmodium vivax using Pvcs and Pvmsp1[J]. Malar J, 2005, 4: 20.
[18]	Choi YK, Choi KM, Park MH, et al. Rapid dissemination of newly introduced Plasmodium vivax genotypes in South Korea[J]. Am J Trop Med Hyg, 2010, 82(3): 426-432.
[19]	Kim JR, Imwong M, Nandy A, et al. Genetic diversity of Plasmodium vivax in Kolkata, India[J]. Malar J, 2006, 5: 71.
[20]	Koepfli C, Mueller I, Marfurt J, et al. Evaluation of Plasmodium vivax genotyping markers for molecular monitoring in clinical trials[J]. J Infect Dis, 2009, 199(7): 1074-1080.
[21]	Kim JR, Nandy A, Maji AK, et al. Genotyping of Plasmodium vivax reveals both short and long latency relapse patterns in Kolkata[J]. PLoS One, 2012, 7(7): e39645.
[22]	Maestre A, Sunil S, Ahmad G, et al. Inter-allelic recombination in the Plasmodium vivax merozoite surface protein 1 gene among Indian and Colombian isolates[J]. Malar J, 2004, 3: 4.
[23]	Khatoon L, Baliraine FN, Bonizzoni M, et al. Genetic structure of Plasmodium vivax and Plasmodium falciparum in the Bannu district of Pakistan[J]. Malar J, 2010, 9: 112.
[24]	Huang B, Huang S, Su XZ, et al. Genetic diversity of Plasmodium vivax population in Anhui Province of China[J]. Malar J, 2014, 13: 13.
[25]	Li YC, Wang GZ, Meng F, et al. Genetic diversity of Plasmodium vivax population before elimination of malaria in Hainan Province, China[J]. Malar J, 2015, 14: 78.
[26]	江钢锋, 李娟兰, 吴少廷, 等. 我国华南间日疟原虫裂殖子表面蛋白1等位基因分型和序列分析[J]. 中国人兽共患病杂志, 2002, 18(2): 57-60.
[27]	耿英芝, 毛玲玲, 腾聪, 等. 辽宁丹东间日疟原虫裂殖子表面蛋白1等位基因型分析[J]. 中国寄生虫学与寄生虫病杂志, 2012, 30(3): 242-244.
[28]	Koepfli C, Rodrigues PT, Antao T, et al. Plasmodium vivax diversity and population structure across four continents[J]. PLoS Negl Trop Dis, 2015, 9(6): e0003872.
[29]	中华人民共和国卫生部, 中华人民共和国卫生行业标准: 疟疾诊断标准(WS259-2006)[S]. 北京: 人民卫生出版社, 2006.
[30]	朱垚吉, 邓艳, 毛祥华, 等. 不同感染来源疟原虫虫株的18S sRNA基因同源性分析[J]. 中国病原生物学杂志, 2014, 9(4): 331-334.
[31]	Putaporntip C, Jongwutiwes S, Sakihama N, et al. Mosaic organization and heterogeneity in frequency of allelic recombination of the Plasmodium vivax merozoite surface protein-1 locus[J]. Proc Natl Acad Sci USA, 2002, 99(25): 16348-16353.
[32]	Anderson TJ, Haubold B, Williams JT, et al. Microsatellite markers reveal a spectrum of population structures in the malaria parasite Plasmodium falciparum[J]. Mol Biol Evol, 2000, 17(10): 1467-1482.
[33]	Soare IS, Barnwel JW, Ferreir MU, et al. A Plasmodium vivax vaccine candidate displays limited allele polymorphism, which does not restrict recognition by antibodies[J]. Mol Med, 1999, (7): 459-470.
[34]	Maneerattanasak S, Gosi P, Krudsood S, et al. Genetic diversity among Plasmodium vivax isolates along the Thai-Myanmar border of Thailand[J]. Malar J, 2016, 15: 75.
[35]	González-Cerón L, Martinez-Barnetche J, Montero-Solís C, et al. Molecular epidemiology of Plasmodium vivax in Latin America: polymorphism and evolutionary relationships of the circumsporozoite gene[J]. Malar J, 2013, 12: 243.
[36]	Zhong D, Bonizzoni M, Zhou G, et al. Genetic diversity of Plasmodium vivax malaria in China and Myanmar[J]. Infect Genet Evol, 2011, 11(6): 1419-1425.
[37]	Putaporntip C, Jongwutiwes S, Tanabe K, et al. Interallelic recombination in the merozoite surface protein 1(MSP-1) gene of Plasmodium vivax from Thai isolates[J]. Mol Biochem Parasitol, 1997, 84(1): 49-56.
[38]	张山鹰, 陆惠民, 许龙善, 等. 我国不同疟区间日疟原虫裂殖子表面蛋白1(PvMSP-1)基因多态性研究[J]. 中国人兽共患病杂志, 2004, 20(1): 26-30.

特征 Characteristics	例数 No. cases	构成比/% Constituent ratio/%	Sal-1基因型 Sal-1 genotype	Belem 基因型 Belem genotype	Recombine基因型 Recombine genotype
性别 Gender
男性 Male	186	90.3	109	45	32
女性 Female	20	9.7	13	3	4
年龄 Age
≤18	15	7.3	7	5	3
19~55	189	90.3	113	43	33
≥56	2	2.4	2	0	0
文化程度 Education
小学及以下 Primary school and below	72	35.0	42	19	11
初中 Middle school	111	53.9	64	25	22
高中及以上 High school and above	23	11.1	16	4	3
职业 Occupation
农民 Farmer	127	61.7	75	32	20
民工 Migrant laborer	47	22.8	27	10	10
其他 Others	32	15.5	20	6	6
既往疟史History of malaria infection
有With	47	22.8	30	9	8
无Without	159	77.2	92	39	28
感染来源地 Source of infection
中国云南省 Yunnan Province,China	14	6.8	7	4	3
缅甸Myanmar	177	86.0	104	42	31
非洲国家 Africa countries	15	7.2	11	2	2

基因型 Genotype	等位基因数 No. alleles	云南感染病例 Cases fromYunnan		缅甸输入病例 Cases imported from Myanmar		非洲输入病例 Cases imported from Africa
基因型 Genotype	等位基因数 No. alleles	等位基因数 No. alleles	期望杂合度 Expected heterozygosity	等位基因数 No. alleles	期望杂合度Expected heterozygosity	等位基因数 No. alleles	期望杂合度Expected heterozygosity
Sal-1	51	9	0.470	50	0.293	10	0.291
Belem	9	2	0.667	9	0.242	2	1.000
Recombine	6	1	-	6	0.373	2	1.000
合计 Total	66	12	-	64	-	14	-