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缅甸金矿流动工人中恶性疟原虫青蒿素耐药性的分子监测

Molecular surveillance of artemisinin resistance falciparum malaria among migrant goldmine workers in Myanmar.

作者信息

Nyunt Myat Htut, Wang Bo, Aye Khin Myo, Aye Kyin Hla, Han Jin-Hee, Lee Seong-Kyun, Han Kay Thwe, Htut Ye, Han Eun-Taek

机构信息

Department of Medical Environmental Biology and Tropical Medicine, School of Medicine, Kangwon National University, Chuncheon, Republic of Korea.

Department of Medical Research, Yangon, Myanmar.

出版信息

Malar J. 2017 Mar 1;16(1):97. doi: 10.1186/s12936-017-1753-8.

Abstract

BACKGROUND

Artemisinin resistance has been reported in Greater Mekong Sub-region countries, including Myanmar. After discovery of artemisinin resistance marker (K13), molecular surveillance on artemisinin resistance in endemic regions have been conducted. As the migrant population represents a high percentage of malaria cases, molecular surveillance of artemisinin resistance among migrant workers is of great concern.

METHODS

A cross-sectional survey was conducted in Shwegyin Township, where migrants work in the goldmines. Blood samples were collected from uncomplicated Plasmodium falciparum-infected migrant workers by active and passive cases screening with rapid diagnostic testing (RDT) and microscopy. Amplification and sequence analysis of artemisinin resistance molecular markers, such as k13, pfarps10, pffd, pfmdr2, pfmrp1, pfrad5, and pfcnbp, were carried out and pfmdr1 copy number analysis was conducted by real-time PCR.

RESULTS

Among the 100 falciparum-infected patients, most were male (90%), of working age (20-40 years) with median parasite density of 11,166 parasites/µL (range 270-110,472 parasites/µL). Artemisinin resistance molecular marker, k13 mutations were detected in (21/100, 21.0%) in which composed of a validated marker, C580Y (9/21, 42.9%) and candidate markers such as P574L (5/21, 23.8%), P667T (5/21, 23.8%) and M476I (2/21, 9.5%). Underlying genetic markers predisposing to become k13 mutants were found as V127M of pfarps10 (41/100, 41.0%), D153Y of pffd (64/100, 64.0%), T484I of pfmdr2 (58/100, 58.0%) and F1390I of pfmrp1 (24/100, 24.0%). The pfmdr1 copy number analysis revealed six copy numbers (1/100, 1.0%), three (2/100, 2.0%), two (8/100, 8.0%) and only one copy number (89/100, 89.0%). Only one sample showed both k13 mutation (P667T) and multiple copy number of pfmdr1.

CONCLUSIONS

High mutant rate of artemisinin resistance markers and relatively high pfmdr1 copy number among isolates collected from migrant goldmine workers alert the importance of containment measures among this target population. Clinical and molecular surveillance of artemisinin resistance among migrants should be scaled up.

摘要

背景

包括缅甸在内的大湄公河次区域国家已报告出现青蒿素耐药性。在发现青蒿素耐药性标志物(K13)后,已在流行地区开展了青蒿素耐药性的分子监测。由于流动人口占疟疾病例的比例很高,因此对农民工中的青蒿素耐药性进行分子监测备受关注。

方法

在瑞茵镇开展了一项横断面调查,该镇有农民工在金矿工作。通过快速诊断检测(RDT)和显微镜检查,对未出现并发症的恶性疟原虫感染农民工进行主动和被动病例筛查,采集血样。对青蒿素耐药性分子标志物,如k13、pfarps10、pffd、pfmdr2、pfmrp1、pfrad5和pfcnbp进行扩增和序列分析,并通过实时PCR进行pfmdr1拷贝数分析。

结果

在100例恶性疟原虫感染患者中,大多数为男性(90%),处于工作年龄(20 - 40岁),寄生虫密度中位数为11,166个寄生虫/微升(范围为270 - 110,472个寄生虫/微升)。在(21/100,21.0%)的患者中检测到青蒿素耐药性分子标志物k13突变,其中包括一个已验证的标志物C580Y(9/21,42.9%)和候选标志物,如P574L(5/21,23.8%)、P667T(5/21,23.8%)和M476I(2/21,9.5%)。发现易成为k13突变体的潜在遗传标志物为pfarps10的V127M(41/100,41.0%)、pffd的D153Y(64/100,64.0%)、pfmdr2的T484I(58/100,58.0%)和pfmrp1的F1390I(24/100,24.0%)。pfmdr1拷贝数分析显示有六个拷贝数(1/100,1.0%)、三个拷贝数(2/100,2.0%)、两个拷贝数(8/100,8.0%)和仅一个拷贝数(89/100,89.0%)。只有一个样本同时显示k13突变(P667T)和pfmdr1的多拷贝数。

结论

从金矿农民工中分离出的菌株中,青蒿素耐药性标志物的高突变率和相对较高的pfmdr1拷贝数提醒人们对这一目标人群采取遏制措施的重要性。应扩大对移民中青蒿素耐药性的临床和分子监测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9e53/5333451/0158626df01d/12936_2017_1753_Fig1_HTML.jpg

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