从疟疾病例流行区返回土耳其的患者中 pfmdr-1、pfcrt 和 pfk13 基因突变的分子调查。

Molecular survey of pfmdr-1, pfcrt, and pfk13 gene mutations among patients returning from Plasmodium falciparum endemic areas to Turkey.

机构信息

Department of Infectious Diseases and Microbiology, University of Health Sciences, Bakırköy Dr. Sadi Konuk Training and Research Hospital, Istanbul, Turkey.

Department of Infectious Diseases and Clinical Microbiology, Dr. Yaşar Eryılmaz Doğubeyazıt State Hospital, Ağrı, Turkey.

出版信息

Malar J. 2024 Sep 27;23(1):286. doi: 10.1186/s12936-024-05107-6.

DOI:10.1186/s12936-024-05107-6

PMID:39334180

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11437951/

Abstract

BACKGROUND

In recent years, there has been an increasing trend in the number of imported Plasmodium falciparum cases in Turkey. To improve treatment success and to better understand malaria epidemiology among imported cases, it is necessary to determine anti-malarial drug resistance. This study aimed to survey polymorphisms of resistance genes in imported P. falciparum patients using archived thin smear preparations and EDTA blood samples.

METHODS

A total of 100 imported P. falciparum patients admitted to Bakırköy Dr. Sadi Konuk Research and Training Hospital between 2017 and 2022 were included in this study. DNA extraction was performed using an archived slide and EDTA blood samples that were microscopically diagnosed. After confirming the samples by real-time PCR, the pfmdr1, pfcrt, and pfk13 genes were amplified and sequenced. Single nucleotide polymorphisms (SNPs) were screened using Geneious R9 software, with the reference P. falciparum clone 3D7 isolate.

RESULTS

All studied samples were confirmed to be P. falciparum using real-time PCR. Nested PCR was conducted and the pfcrt (92 samples), pfmdr1 (91 samples), and pfk13 (93 samples) genes were successfully amplified. Sequence analysis revealed the highest mutation rate in the pfmdr1 (74.5%) gene, with the identification of five haplotypes: NYSND (wild-type, 23%), NFSND (56%), NYSDD (2.2%), NFSDD (15.4%), and YFSND (3.4%)]. The pfcrt mutation was identified in 11 samples (12.2%), whereas the pfk13 mutation was found in only two samples.

CONCLUSION

This study is the first molecular survey of anti-malarial drug resistance genes in Turkey. With the increasing number of imported Plasmodium malaria cases and recent reports of sporadic indigenous P. falciparum cases, malaria is becoming a growing concern in Turkey. Although molecular screening for resistance markers in P. falciparum malaria is not routinely conducted, the data from this study will enhance treatment success rates and contribute to global malaria elimination.

摘要

背景

近年来，土耳其输入性疟疾病例呈上升趋势。为提高治疗成功率，更好地了解输入性疟疾病例的流行病学情况，有必要确定抗疟药物的耐药性。本研究旨在利用存档的薄血涂片和 EDTA 血样检测输入性恶性疟原虫患者的耐药基因的多态性。

方法

本研究共纳入 2017 年至 2022 年期间入住 Bakırköy Dr. Sadi Konuk 研究与培训医院的 100 例输入性恶性疟原虫患者。采用存档的玻片和显微镜诊断的 EDTA 血样提取 DNA。使用实时 PCR 确认样本后，扩增和测序 pfmdr1、pfcrt 和 pfk13 基因。使用 Geneious R9 软件筛选单核苷酸多态性（SNP），以恶性疟原虫克隆 3D7 分离株作为参考。

结果

所有研究样本均通过实时 PCR 确认为恶性疟原虫。进行巢式 PCR，成功扩增 pfcrt（92 个样本）、pfmdr1（91 个样本）和 pfk13（93 个样本）基因。序列分析显示 pfmdr1 基因的突变率最高（74.5%），鉴定出 5 种单倍型：NYSND（野生型，23%）、NFSND（56%）、NYSDD（2.2%）、NFSDD（15.4%）和 YFSND（3.4%）。pfcrt 突变发生在 11 个样本（12.2%）中，而 pfk13 突变仅发生在 2 个样本中。