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乌干达北部成年有症状患者分离出的恶性疟原虫中K13螺旋桨基因多态性的流行情况。

Prevalence of K13-propeller gene polymorphisms among Plasmodium falciparum parasites isolated from adult symptomatic patients in northern Uganda.

作者信息

Ocan Moses, Bwanga Freddie, Okeng Alfred, Katabazi Fred, Kigozi Edgar, Kyobe Samuel, Ogwal-Okeng Jasper, Obua Celestino

机构信息

Department of Pharmacology & Therapeutics, Makerere University, P. O. Box 7072, Kampala, Uganda.

Department of Medical Microbiology, Makerere University, P. O. Box 7072, Kampala, Uganda.

出版信息

BMC Infect Dis. 2016 Aug 19;16(1):428. doi: 10.1186/s12879-016-1777-7.

Abstract

BACKGROUND

In the absence of an effective vaccine, malaria treatment and eradication is still a challenge in most endemic areas globally. This is especially the case with the current reported emergence of resistance to artemisinin agents in Southeast Asia. This study therefore explored the prevalence of K13-propeller gene polymorphisms among Plasmodium falciparum parasites in northern Uganda.

METHODS

Adult patients (≥18 years) presenting to out-patients department of Lira and Gulu regional referral hospitals in northern Uganda were randomly recruited. Laboratory investigation for presence of plasmodium infection among patients was done using Plasmodium falciparum exclusive rapid diagnostic test, histidine rich protein-2 (HRP2) (Pf). Finger prick capillary blood from patients with a positive malaria test was spotted on a filter paper Whatman no. 903. The parasite DNA was extracted using chelex resin method and sequenced for mutations in K13-propeller gene using Sanger sequencing. PCR DNA sequence products were analyzed using in DNAsp 5.10.01software, data was further processed in Excel spreadsheet 2007.

RESULTS

A total of 60 parasite DNA samples were sequenced. Polymorphisms in the K13-propeller gene were detected in four (4) of the 60 parasite DNA samples sequenced. A non-synonymous polymorphism at codon 533 previously detected in Cambodia was found in the parasite DNA samples analyzed. Polymorphisms at codon 522 (non-synonymous) and codon 509 (synonymous) were also found in the samples analyzed. The study found evidence of positive selection in the Plasmodium falciparum population in northern Uganda (Tajima's D = -1.83205; Fu and Li's D = -1.82458).

CONCLUSIONS

Polymorphism in the K13-propeller gene previously reported in Cambodia has been found in the Ugandan Plasmodium falciparum parasites. There is need for continuous surveillance for artemisinin resistance gene markers in the country.

摘要

背景

在缺乏有效疫苗的情况下,疟疾治疗和根除在全球大多数流行地区仍是一项挑战。东南亚目前报告出现对青蒿素类药物的耐药性,情况尤其如此。因此,本研究探讨了乌干达北部恶性疟原虫寄生虫中K13螺旋桨基因多态性的流行情况。

方法

随机招募到乌干达北部Lira和Gulu地区转诊医院门诊部就诊的成年患者(≥18岁)。使用恶性疟原虫专用快速诊断试验、富含组氨酸蛋白-2(HRP2)(Pf)对患者进行疟原虫感染的实验室检测。将疟疾检测呈阳性患者的手指刺血毛细血管血点在Whatman No. 903滤纸上。使用螯合树脂法提取寄生虫DNA,并使用桑格测序法对K13螺旋桨基因中的突变进行测序。使用DNAsp 5.10.01软件分析PCR DNA序列产物,数据在Excel 2007电子表格中进一步处理。

结果

共对60个寄生虫DNA样本进行了测序。在测序的60个寄生虫DNA样本中,有4个检测到K13螺旋桨基因多态性。在分析的寄生虫DNA样本中发现了先前在柬埔寨检测到的第533位密码子的非同义多态性。在分析的样本中还发现了第522位密码子(非同义)和第509位密码子(同义)的多态性。该研究发现乌干达北部恶性疟原虫种群存在正选择的证据( Tajima's D = -1.83205;Fu和Li's D = -1.82458)。

结论

在乌干达恶性疟原虫寄生虫中发现了先前在柬埔寨报告的K13螺旋桨基因多态性。该国需要持续监测青蒿素耐药基因标记。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f18f/4992308/cff60d98cb40/12879_2016_1777_Fig1_HTML.jpg

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