Analysis of genetic polymorphisms of merozoite surface protein-1 and circumsporozoite protein of Plasmodium vivax

doi:10.12140/j.issn.1000-7423.2020.03.011

Abstract

Abstract:

Objective This study aimed to conduct descriptive analysis of the genetic polymorphisms of Plasmodium vivax gene merozoite surface protein-1 (PvMSP-1) and circumsporozoite protein (PvCSP) in vivax malaria cases in Hangzhou City. Methods Blood samples and epidemiological information of vivax malaria cases reported in Hangzhou during 2008-2019 were collected. P. vivax genomic DNA was extracted by QIAamp DNA blood mini kit. Pvmsp-1 and Pvcsp were amplified by nested PCR and the amplification products were sequenced. The amino-acid sequences were analyzed using DNAStar and MEGA 6.0 softwares and the phylogenetic tree was constructed using the neighbor-joining method. The descriptive and inductive analysis of genetic polymorphism and place of infection origin were performed using SPSS 17.0. Results Fifty-six vivax malaria cases were sampled, of whom 44 and 54 samples were successfully sequenced for Pvmsp-1 and Pvcsp, respectively. Sequence analysis indicated that 44 Pvmsp-1 sequences were clustered into four genotypes: SalⅠ(18 samples), Belem (8), R-Ⅲ (17) and R-Ⅳ (1); 54 Pvcsp sequences were devided into two genotypes: VK210 (52 samples) and VK247 (2). Analysis of the genotyping and place of infection origin of Pvmsp-1 gene showed that the SalⅠtype was widely distributed in East China, Southeast Asia, India, and Ethiopia. The main type of Pvmsp-1 in East China was R-Ⅲ. In addition, the R-Ⅲ type was also distributed in South Asia and Southeast Asia. The Belem type was mainly distributed in Southeast Asia, South Asia and Ethiopia. Analysis of Pvcsp gene polymorphism and place of infection origin revealed four types of polymorphism with some regional features. Conclusion The Pvmsp-1 and Pvcsp genes in vivax malaria cases reported in Hangzhou during 2008-2019 were characterized being highly polymorphic, with some genotypes having certain regional features.

Key words: Plasmodium vivax, Merozoite surface protein-1, Circumsporozoite protein, Genetic diversity

CLC Number:

R531.31

JIN Hang-yi, ZHANG Ling-ling, ZHU Su-juan, XU Wei-min, CHEN Jun-fang, RUAN Wei, YAO Li-nong, CHEN Hua-liang. Analysis of genetic polymorphisms of merozoite surface protein-1 and circumsporozoite protein of Plasmodium vivax[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2020, 38(3): 323-331.

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基因 Genes	引物名称 Primer names	引物序列（5′→3′） Primer sequences (5′→3′)
Pvmsp-1	Pvmsp-1-F1	CCCTACTACTTGATGGTCCTC
	Pvmsp-1-R1	CCTTCTGGTACAGCTCAATG
	Pvmsp-1-F2	AGCATGATCGCCACTGAGAAG
	Pvmsp-1-R2	GTGCTTGTGACATGCGTAAGC
Pvcsp	VCSF1	ATGTAGATCTGTCCAAGGCCATAAA
	VCSR1	TAATTGAATAATGCTAGGACTAACAATATG
	VCSF2	GCAGAACCAAAAAATCCACGTGAAAATAAG
	VCSR2	CCAACGGTAGCTCTAACTTTATCTAGGTAT

来源地 Source area	Pvmsp-1/Pvcsp												合计Total
来源地 Source area	2008	2009	2010	2011	2012	2013	2014	2015	2016	2017	2018	2019	合计Total
中国安徽Anhui, China	1, 2	0, 0	4, 4	4, 4	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	9, 10
中国浙江Zhejiang, China	0, 0	0, 0	0, 0	1, 1	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	1, 1
中国四川Sichuan, China	0, 1	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 1
柬埔寨Cambodia	0, 0	0, 0	0, 0	2, 2	2, 3	2, 2	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	6, 7
缅甸Myanmar	0, 0	0, 0	0, 0	0, 1	0, 1	0, 0	0, 0	2, 2	0, 0	0, 0	0, 0	0, 0	2, 4
印度尼西亚Indonesia	0, 0	0, 0	0, 0	0, 0	1, 1	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	1, 1
印度India	0, 0	0, 0	0, 0	2, 2	0, 0	0, 0	1, 1	0, 0	0, 0	1, 1	0, 0	0, 0	4, 4
巴基斯坦Pakistan	0, 0	0, 0	0, 0	2, 3	0, 0	1, 2	0, 0	1, 1	1, 3	1, 1	0, 0	1, 1	7, 11
阿富汗Afghanistan	0, 0	0, 0	0, 0	0, 0	0, 0	2, 2	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	2, 2
埃塞俄比亚Ethiopia	0, 0	0, 0	0, 0	0, 1	0, 0	0, 0	0, 0	2, 2	0, 0	3, 3	0, 0	0, 0	5, 6
尼日利亚Nigeria	0, 0	1, 1	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	1, 1	2, 2
苏丹Sudan	0, 0	0, 0	0, 0	0, 0	0, 0	1, 1	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	1, 1
科特迪瓦Cote d′ivoire	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 1	0, 0	0, 1
非洲（不详）Africa	0, 0	1, 1	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	1, 1
未知来源地Unknown	0, 0	0, 0	2, 1	1, 1	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	0, 0	3, 2
合计Total	1, 3	2, 2	6, 5	12, 15	3, 5	6, 7	1, 1	5, 5	1, 3	5, 5	0, 1	2, 2	44, 54

来源地Area	V1				V2		V3				V4						V5
来源地Area	a	b	c	d	e	f	g	h	i	j	k	l	m	n	o	p	q	r	s	t	u	v	w
中国安徽Anhui, China	1	0	8	0	10	0	0	10	0	0	0	1	0	3	4	2	0	2	0	1	3	4	0
中国浙江Zhejiang, China	0	0	1	0	1	0	0	1	0	0	0	1	0	0	0	0	0	0	0	0	0	0	1
中国四川Sichuan, China	0	0	0	0	1	0	1	0	0	0	0	1	0	0	0	0	0	1	0	0	0	0	0
柬埔寨Cambodia	3	1	2	0	5	2	4	1	1	1	0	2	4	1	0	0	1	5	1	0	0	0	0
缅甸Myanmar	2	0	0	0	1	1	0	1	1	0	0	1	0	1	0	0	0	1	1	0	0	0	0
印度尼西亚Indonesia	0	1	0	0	0	1	0	0	1	0	0	0	0	1	0	0	0	1	0	0	0	0	0
印度India	4	0	0	0	4	0	4	0	0	0	1	3	0	0	0	0	0	4	0	0	0	0	0
巴基斯坦Pakistan	3	1	3	0	11	0	11	0	0	0	0	11	0	0	0	0	0	11	0	0	0	0	0
阿富汗Afghanistan	0	0	2	0	2	0	2	0	0	0	0	2	0	0	0	0	0	2	0	0	0	0	0
埃塞俄比亚Ethiopia	1	3	1	0	6	0	6	0	0	0	0	6	0	0	0	0	0	6	0	0	0	0	0
尼日利亚Nigeria	1	1	0	0	2	0	2	0	0	0	0	2	0	0	0	0	0	2	0	0	0	0	0
苏丹Sudan	1	0	0	0	1	0	1	0	0	0	0	1	0	0	0	0	0	1	0	0	0	0	0
科特迪瓦Cote d’ivoire	0	0	0	0	1	0	1	0	0	0	0	1	0	0	0	0	0	1	0	0	0	0	0
非洲（不详）Africa	0	0	0	1	1	0	1	0	0	0	0	1	0	0	0	0	0	1	0	0	0	0	0
未知Unknown	2	1	0	0	2	0	1	1	0	0	0	1	0	0	0	1	0	1	0	0	1	0	0

基因 Gene	多态性特征 Polymorphism characteristics	地域分布 Regional distribution
基因 Gene	多态性特征 Polymorphism characteristics	中国华东地区 East China	东南亚地区 Southeast Asia	南亚地区 South Asia	非洲地区 Africa
Pvmsp-1	SalⅠ	√	√	√	√
	Belem		√	√	√
	R-Ⅲ	√	√	√	√
	R-Ⅳ				√
Pvcsp	中央重复区后不具有12肽插入序列 Without a 12-peptide insertion sequence after the central repeat region		√
	12肽插入序列后GGNA重复次数 Number of repeats of the GGNA unit after the 12-peptide insertion sequence	2	1~4	1	1
	中央重复区9肽单位重复次数 Number of repeats of peptide repeat motif in central repeat region	18~22	18~20	18	18
	9肽单位（GNGAGGQAA）重复次数 Number of repeats of peptide repeat motif (GNGAGGQAA)	1, 13~15	1~2	1	1