云南省恶性疟原虫K13基因Kelch结构域与青蒿素耐药相关的多态性分析

Ai-ming Sun, Ying Dong, Meng-ni Chen, Yan-chun Xu, Yan Deng, Xiang-hua Mao, Jian Wang
{"title":"云南省恶性疟原虫K13基因Kelch结构域与青蒿素耐药相关的多态性分析","authors":"Ai-ming Sun,&nbsp;Ying Dong,&nbsp;Meng-ni Chen,&nbsp;Yan-chun Xu,&nbsp;Yan Deng,&nbsp;Xiang-hua Mao,&nbsp;Jian Wang","doi":"","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To investigate the polymorphism of Plasmodium falciparum K13 gene kelch domain region and provide basis for understanding the artemisinin resistance of falciparum malaria in Yunnan Province.</p><p><strong>Methods: </strong>The filter blood samples and relative information of falciparum malaria cases were collected in 16 prefectures of Yunnan Province from January 2013 to December 2015. The source of infection was determined by epidemiological investigation and the place of case discovery was confirmed according to the China Information System for Disease Control and Prevention Epidemic Registration. The K13 gene kelch domain region was amplified by nested PCR, sequenced, and blasted against the reference strain 3D7(PF3D7_1343700). The K13 gene kelch domain region polymorphism was analyzed with Mega 5.04. The variable sites and genetic distance between sequences were analyzed. The constituent ratio of amino acid mutation sites was calculated and analyzed with χ2 test.</p><p><strong>Results: </strong>A total of 202 blood samples were collected from 2013 to 2015, comprising 190 from imported cases, 12 from local cases in Yunnan Province. The constitutent ratio of infection cases were 30.7% (62/202), 34.2% (69/202) and 35.1% (71/202) respectively, increased year by year. The K13 gene kelch domain was successfully amplified from 192 samples and 190 were successfully sequenced, detecting missense mutation of K13 gene in 66 samples, the mutation rate was 34.7% (66/190). The detection rate of K13 gene mutation was 40.9% (27/66), 37.9% (25/66) and 21.2% (14/66) respectively, decreased year by year. In this study, ten types of mutations were detected, which were F446I, A578S, N458Y, P574L, A676D, G449A, C469Y, V494I, E556D and S16L. The highest mutation rate occurred in F446I which was 72.7% (48/66). The proportion of F446I mutation type was 58.3% (28/48) in an age-range of 18-56 years, 70.8% (34/48) in farmers, and 91.7% (44/48) in patients with infection source in Southeast Asia, all significantly higher than that of other groups with the same characteristics (41.7%, 20/48; 29.2%, 14/48; and 8.3%, 4/48, respectively)(χ2=4.633, 5.556 and 5.152, both P<0.05). There was a 248 bp homologous sequence in the 190 sequences, composed of 235 conservative sites (94.8%), 13 variable sites (5.2%), 5 parsim-info sites (2.0%), and 8 singleton sites (3.2%). The genetic distance among the 190 sequences ranged 0.000-0.036, with an average of 0.001±0.001.</p><p><strong>Conclusion: </strong>There are 10 types of mutations in the K13 kelch domain in Yunnan Province, the predominant mutation type was F446I.</p>","PeriodicalId":23981,"journal":{"name":"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases","volume":"34 4","pages":"339-45"},"PeriodicalIF":0.0000,"publicationDate":"2016-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[Polymorphism Analysis of Plasmodium falciparum K13 Gene Kelch Domain Associated with Resistance to Artemisinin in Yunnan Province].\",\"authors\":\"Ai-ming Sun,&nbsp;Ying Dong,&nbsp;Meng-ni Chen,&nbsp;Yan-chun Xu,&nbsp;Yan Deng,&nbsp;Xiang-hua Mao,&nbsp;Jian Wang\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>To investigate the polymorphism of Plasmodium falciparum K13 gene kelch domain region and provide basis for understanding the artemisinin resistance of falciparum malaria in Yunnan Province.</p><p><strong>Methods: </strong>The filter blood samples and relative information of falciparum malaria cases were collected in 16 prefectures of Yunnan Province from January 2013 to December 2015. The source of infection was determined by epidemiological investigation and the place of case discovery was confirmed according to the China Information System for Disease Control and Prevention Epidemic Registration. The K13 gene kelch domain region was amplified by nested PCR, sequenced, and blasted against the reference strain 3D7(PF3D7_1343700). The K13 gene kelch domain region polymorphism was analyzed with Mega 5.04. The variable sites and genetic distance between sequences were analyzed. The constituent ratio of amino acid mutation sites was calculated and analyzed with χ2 test.</p><p><strong>Results: </strong>A total of 202 blood samples were collected from 2013 to 2015, comprising 190 from imported cases, 12 from local cases in Yunnan Province. The constitutent ratio of infection cases were 30.7% (62/202), 34.2% (69/202) and 35.1% (71/202) respectively, increased year by year. The K13 gene kelch domain was successfully amplified from 192 samples and 190 were successfully sequenced, detecting missense mutation of K13 gene in 66 samples, the mutation rate was 34.7% (66/190). The detection rate of K13 gene mutation was 40.9% (27/66), 37.9% (25/66) and 21.2% (14/66) respectively, decreased year by year. In this study, ten types of mutations were detected, which were F446I, A578S, N458Y, P574L, A676D, G449A, C469Y, V494I, E556D and S16L. The highest mutation rate occurred in F446I which was 72.7% (48/66). The proportion of F446I mutation type was 58.3% (28/48) in an age-range of 18-56 years, 70.8% (34/48) in farmers, and 91.7% (44/48) in patients with infection source in Southeast Asia, all significantly higher than that of other groups with the same characteristics (41.7%, 20/48; 29.2%, 14/48; and 8.3%, 4/48, respectively)(χ2=4.633, 5.556 and 5.152, both P<0.05). There was a 248 bp homologous sequence in the 190 sequences, composed of 235 conservative sites (94.8%), 13 variable sites (5.2%), 5 parsim-info sites (2.0%), and 8 singleton sites (3.2%). The genetic distance among the 190 sequences ranged 0.000-0.036, with an average of 0.001±0.001.</p><p><strong>Conclusion: </strong>There are 10 types of mutations in the K13 kelch domain in Yunnan Province, the predominant mutation type was F446I.</p>\",\"PeriodicalId\":23981,\"journal\":{\"name\":\"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases\",\"volume\":\"34 4\",\"pages\":\"339-45\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2016-08-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

摘要

目的:研究云南省恶性疟原虫K13基因kelch结构域的多态性,为了解云南省恶性疟疾对青蒿素的耐药性提供依据。方法:采集2013年1月至2015年12月云南省16个地市恶性疟疾病例的过滤血样及相关资料。流行病学调查确定传染源,病例发现地点根据中国疾病预防控制信息系统疫情登记确认。采用巢式PCR扩增K13基因kelch结构域,对其进行测序,并与参考菌株3D7(PF3D7_1343700)进行blast。利用Mega 5.04分析K13基因kelch结构域多态性。分析了变异位点和序列间的遗传距离。氨基酸突变位点组成比采用χ2检验进行统计分析。结果:2013 - 2015年共采集标本202份,其中输入病例190份,本地病例12份。感染病例构成比分别为30.7%(62/202)、34.2%(69/202)和35.1%(71/202),呈逐年上升趋势。从192份样本中成功扩增出K13基因kelch结构域,并对190份样本进行测序,其中66份样本检测到K13基因错义突变,突变率为34.7%(66/190)。K13基因突变检出率分别为40.9%(27/66)、37.9%(25/66)和21.2%(14/66),呈逐年下降趋势。本研究共检测到10种突变类型,分别为F446I、A578S、N458Y、P574L、A676D、G449A、C469Y、V494I、E556D和S16L。F446I的突变率最高,为72.7%(48/66)。F446I突变型在18-56岁人群中占58.3%(28/48),在农民中占70.8%(34/48),在东南亚感染源患者中占91.7%(44/48),均显著高于其他相同特征人群(41.7%,20/48;29.2%, 14/48;8.3%, 4/48)(χ2=4.633、5.556、5.152,P均<0.05)。190个序列中同源序列为248 bp,由235个保守位点(94.8%)、13个可变位点(5.2%)、5个parsim-info位点(2.0%)和8个单点位点(3.2%)组成。190个序列的遗传距离在0.000 ~ 0.036之间,平均为0.001±0.001。结论:云南地区K13 kelch结构域存在10种突变类型,以F446I突变类型为主。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Polymorphism Analysis of Plasmodium falciparum K13 Gene Kelch Domain Associated with Resistance to Artemisinin in Yunnan Province].

Objective: To investigate the polymorphism of Plasmodium falciparum K13 gene kelch domain region and provide basis for understanding the artemisinin resistance of falciparum malaria in Yunnan Province.

Methods: The filter blood samples and relative information of falciparum malaria cases were collected in 16 prefectures of Yunnan Province from January 2013 to December 2015. The source of infection was determined by epidemiological investigation and the place of case discovery was confirmed according to the China Information System for Disease Control and Prevention Epidemic Registration. The K13 gene kelch domain region was amplified by nested PCR, sequenced, and blasted against the reference strain 3D7(PF3D7_1343700). The K13 gene kelch domain region polymorphism was analyzed with Mega 5.04. The variable sites and genetic distance between sequences were analyzed. The constituent ratio of amino acid mutation sites was calculated and analyzed with χ2 test.

Results: A total of 202 blood samples were collected from 2013 to 2015, comprising 190 from imported cases, 12 from local cases in Yunnan Province. The constitutent ratio of infection cases were 30.7% (62/202), 34.2% (69/202) and 35.1% (71/202) respectively, increased year by year. The K13 gene kelch domain was successfully amplified from 192 samples and 190 were successfully sequenced, detecting missense mutation of K13 gene in 66 samples, the mutation rate was 34.7% (66/190). The detection rate of K13 gene mutation was 40.9% (27/66), 37.9% (25/66) and 21.2% (14/66) respectively, decreased year by year. In this study, ten types of mutations were detected, which were F446I, A578S, N458Y, P574L, A676D, G449A, C469Y, V494I, E556D and S16L. The highest mutation rate occurred in F446I which was 72.7% (48/66). The proportion of F446I mutation type was 58.3% (28/48) in an age-range of 18-56 years, 70.8% (34/48) in farmers, and 91.7% (44/48) in patients with infection source in Southeast Asia, all significantly higher than that of other groups with the same characteristics (41.7%, 20/48; 29.2%, 14/48; and 8.3%, 4/48, respectively)(χ2=4.633, 5.556 and 5.152, both P<0.05). There was a 248 bp homologous sequence in the 190 sequences, composed of 235 conservative sites (94.8%), 13 variable sites (5.2%), 5 parsim-info sites (2.0%), and 8 singleton sites (3.2%). The genetic distance among the 190 sequences ranged 0.000-0.036, with an average of 0.001±0.001.

Conclusion: There are 10 types of mutations in the K13 kelch domain in Yunnan Province, the predominant mutation type was F446I.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信