中国血吸虫病防治杂志 ›› 2020, Vol. 32 ›› Issue (6): 612-.

• 实验研究 • 上一篇    下一篇

2015—2016年山东省由赤道几内亚输入的恶性疟原虫抗药性基因多态性分析

聂广馗1, 2, 3，徐超1*，魏庆宽1, 2，李瑾1，肖婷1，孙慧1, 2，孔祥礼1，尹昆1, 2，赵桂华1，张本光1，闫歌1，黄炳成1, 2*   

  1. 1 山东省寄生虫病防治研究所、山东第一医科大学（山东省医学科学院）（济宁 272033）；2 济南大学、山东省医学科学院医学与生命科学学院；3山东省济宁卫生学校
  • 出版日期:2020-12-08 发布日期:2020-12-08
  • 作者简介:聂广馗，男，讲师。研究方向：寄生虫分子生物学
  • 基金资助:
    山东省自然科学基金（ZR2014YL036、ZR2017YL003、ZR2017YL005）；山东省医药卫生科技发展计划（2016WS0394）；山东省医学科学院课题计划（2017?42）；山东省医学科学院医药卫生科技创新工程；山东第一医科大学“学术提升”计划（2019QL005)

Analysis of drug-resistant gene polymorphisms in Plasmodium falciparum imported from Equatorial Guinea to Shandong Province in 2015 and 2016

NIE Guang-Kui1, 2, 3, XU Chao1*, WEI Qing-Kuan1, 2, LI Jin1, XIAO Ting1, SUN Hui1, 2, KONG Xiang-Li1, YIN Kun1,2, ZHAO Gui-Hua1, ZHANG Ben-Guang1, YAN Ge1, HUANG Bing-Cheng1, 2*   

  1. 1 Shandong Institute of Parasitic Diseases, Shandong First Medical University & Shandong Academy of Medical Sciences, Jining 272033, China； 2 School of Medicine and Life Sciences, Shandong Academy of Medical Sciences, University of Jinan, China； 3 Jining Health School, Shandong Province, China
  • Online:2020-12-08 Published:2020-12-08

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摘要： 目的　了解山东省由赤道几内亚输入的恶性疟原虫抗性基因多态性情况。方法　采集2015—2016年山东省由赤道几内亚务工返乡的输入性恶性疟患者血样，提取疟原虫基因组DNA，对恶性疟原虫抗性基因Pfcrt、Pfmdr1、Pfdhfr、Pfdhps、K13进行套式PCR扩增、DNA测序和序列对比分析。结果　全部样本5种抗性基因目的片段均成功扩增和测序。Pfcrt野生型、突变型、混合型分别占72.8%、18.6%、8.6%，突变型全部为CVIET（下划线表示突变位点，下同）；Pfmdr1野生型、突变型、混合型分别占20.0%、61.4%、18.6%，突变型主要为YF和NF；Pfdhfr野生型、突变型、混合型分别占1.4%、98.6%、0，突变型主要为AIRNI；Pfdhps野生型、突变型、混合型分别占1.4%、94.3%、4.3%，突变型主要为SGKAA；Pfdhfr和Pfdhps完全抗性基因型IRNGE占8.6%；1.4%的样本K13基因发生A578S突变。结论　山东省由赤道几内亚输入的恶性疟原虫Pfcrt、Pfmdr1、Pfdhfr、Pfdhps、K13基因均发生不同程度突变；Pfcrt突变型比例较低，Pfmdr1、Pfdhfr、Pfdhps突变型比例较高；检测到K13基因发生A578S突变。 　

关键词: 恶性疟原虫, 输入性疟疾, 赤道几内亚, 抗药性, 基因多态性

Abstract: Objective　To investigate the drug?resistant gene polymorphisms in Plasmodium falciparum imported from Equatorial Guinea to Shandong Province. Methods　From 2015 to 2016, blood samples were collected from imported P. falciparum malaria patients returning from Equatorial Guinea to Shandong Province, and genome DNA of the malaria parasite was extracted. The drug?resistant Pfcrt, Pfmdr1, Pfdhfr, Pfdhps, and K13 genes of P. falciparum were amplified using a PCR assay, followed by DNA sequencing, and the sequences were aligned. Results The target fragments of all 5 drug?resistant genes of P. falciparum were successfully amplified and sequenced. There were 72.8%, 18.6%, and 8.6% of P. falciparum parasites carrying the wild?, mutant?, and mixed?type Pfcrt gene, respectively, and all mutant haplotypes were CVIET (the underline indicates the mutation site). There were 20.0%, 61.4% and 18.6% of P. falciparum parasites carrying the wild?, mutant?, and mixed?type Pfmdr1 gene, respectively, and the mutant haplotypes mainly included YF and NF (the underlines indicate the mutation sites). There were 1.4%, 98.6%, and 0 of P. falciparum parasites carrying the wild?, mutant?, and mixed?type Pfdhfr gene, respectively, and AIRNI was the predominant mutant haplotype (the underline indicates the mutation site). There were 1.4%, 94.3%, and 4.3% of P. falciparum parasites carrying the wild?, mutant?, and mixed?type Pfdhps gene, respectively, and SGKAA was the predominant mutant haplotype (the underline indicates the mutation site). The complete drug?resistant IRNGE genotype consisted of 8.6% of the Pfdhfr and Pfdhps genes, and the K13 gene A578S mutation occurred in 1.4% of the parasite samples. Conclusion　There are mutations in the Pfcrt, Pfmdr1, Pfdhfr, Pfdhps, and K13 genes of P. falciparum imported from Equatorial Guinea to Shandong Province, with a low frequency in the Pfcrt gene mutation and a high frequency in the Pfmdr1, Pfdhfr, and Pfdhps gene mutations, and the K13 gene A578S mutation is detected in the parasite samples.

Key words: Plasmodium falciparum, Imported malaria, Equatorial Guinea, Drug resistance, Gene polymorphism

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