一种用于检测恶性疟原虫青蒿素抗性的新型表型检测方法-环状体阶段生存测定法(rsa)及其改进的综述

张捷,; Zhang, Jie; 冯国华,; 邹春燕,; 苏品璨,; 刘怀鄂,; 杨照青,; Feng, Guo-Hua; Zou, Chunyan; Su, Peng; Liu, Huaie; Zhou, Yang

doi:10.24272/j.issn.2095-8137.2017.075

Cited by 11 publications

(7 citation statements)

References 26 publications

(36 reference statements)

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“…We also did not find significant correlations in drug sensitivity among quinoline drugs and artemisinin family drugs, suggesting that in the emergence of CQ resistance in P. vivax, ACT could be considered as an option for the treatment of CQ-resistant vivax malaria like the practice in Indonesia [8,9]. Of note, sensitivities to artemisinin family drugs may also need to be measured using an assay similar to the in vitro ring-stage survival assay developed for P. falciparum [69,70].…”

Section: Discussionmentioning

confidence: 83%

Ex vivo susceptibilities of Plasmodium vivax isolates from the China-Myanmar border to antimalarial drugs and association with polymorphisms in Pvmdr1 and Pvcrt-o genes

Zhang

et al. 2020

PLoS Negl Trop Dis

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Background Vivax malaria is an important public health problem in the Greater Mekong Subregion (GMS), including the China-Myanmar border. Previous studies have found that Plasmodium vivax has decreased sensitivity to antimalarial drugs in some areas of the GMS, but the sensitivity of P. vivax to antimalarial drugs is unclear in the China-Myanmar border. Here, we investigate the drug sensitivity profile and genetic variations for two drug resistance related genes in P. vivax isolates to provide baseline information for future drug studies in the China-Myanmar border. Methodology/Principal findings A total of 64 P. vivax clinical isolates collected from the China-Myanmar border area were assessed for ex vivo susceptibility to eight antimalarial drugs by the schizont maturation assay. The medians of IC 50 (half-maximum inhibitory concentrations) for chloroquine, mefloquine, pyronaridine, piperaquine, quinine, artesunate, artemether, dihydroartemisinin were 84.2 nM, 34.9 nM, 4.0 nM, 22.3 nM, 41.4 nM, 2.8 nM, 2.1 nM and 2.0 nM, respectively. Twelve P. vivax clinical isolates were found over the cutoff IC 50 value (220 nM) for chloroquine resistance. In addition, sequence polymorphisms in pvmdr1 (P. vivax multidrug resistance-1), pvcrt-o (P. vivax chloroquine resistance transporter-o), and difference in pvmdr1 copy number were studied. Sequencing of the pvmdr1 gene in 52 samples identified 12 amino acid substitutions, among which two (G698S and T958M) were fixed, M908L were present in 98.1% of the isolates, while Y976F and F1076L were present in 3.8% and 78.8% of the isolates, respectively. Amplification of the pvmdr1 gene was only detected in 4.8% of PLOS NEGLECTED TROPICAL DISEASES

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Section: Discussionmentioning

confidence: 83%

Ex vivo susceptibilities of Plasmodium vivax isolates from the China-Myanmar border to antimalarial drugs and association with polymorphisms in Pvmdr1 and Pvcrt-o genes

Zhang

et al. 2020

PLoS Negl Trop Dis

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“…RSA was performed as previously described (Siddiqui et al, 2018; Witkowski et al, 2013a; Zhang et al, 2017). Briefly, highly synchronous parasite cultures at the young ring stage (0–3 h) were exposed to 700 nM DHA or 0.1% dimethyl sulfoxide (DMSO) as the control for 6 h. The drugs were then washed off with RPMI 1640, and parasites cultivated further for 66 h under the standard in vitro culture conditions.…”

Section: Methodsmentioning

confidence: 99%

In vitro susceptibility of Plasmodium falciparum isolates from the China-Myanmar border area to artemisinins and correlation with K13 mutations

Zhang

Siddiqui

et al. 2019

International Journal for Parasitology: Drugs and Drug Resistan

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Mutations in the Kelch domain of the K13 gene (PF3D7_1343700) were previously associated with artemisinin resistance in Plasmodium falciparum . This study followed the dynamics of the K13 polymorphisms in P. falciparum parasites from the China-Myanmar border area obtained in 2007–2016, and their in vitro sensitivities to artesunate (AS) and dihydroartemisinin (DHA). The 50% effective concentration (EC 50 72h ) values of 133 culture-adapted field isolates to AS and DHA, measured by the conventional 72 h SYBR Green I-based assay, varied significantly among the parasites from different years; all were significantly higher than that of the reference strain 3D7. Compared with parasites from 2007 to 2008, ring survival rates almost doubled in parasites obtained in later years. Sequencing the full-length K13 genes identified 11 point mutations present in 85 (63.9%) parasite isolates. F446I was the predominant (55/133) variant, and its frequency was increased from 17.6% (3/17) in 2007 to 55.9% (19/34) in 2014–2016. No wild-type (WT) Kelch domain sequences were found in the 34 samples obtained from 2014 to 2016. In the 2014–2016 samples, a new mutation (G533S) appeared and reached 44.1% (15/34). Collectively, parasites with the Kelch domain mutations (after amino acid 440) had significantly higher ring survival rates than the WT parasites. Individually, F446I, G533S and A676D showed significantly higher ring survival rates than the WT. Although the drug sensitivity phenotypes measured by the RSA 6h and EC 50 72h assays may be intrinsically linked to the in vivo clinical efficacy data, the values determined by these two assays were not significantly correlated. This study identified the trend of K13 mutations in parasite populations from the China-Myanmar border area, confirmed an overall correlation of Kelch domain mutations with elevated ring-stage survival rates, and emphasized the importance of monitoring the evolution and spread of parasites with reduced artemisinin sensitivity along the malaria elimination course.

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“…Despite a variety of available techniques to quantify parasite biomass, the applied incubation times are usually in the range of 48–72 h, though there are clear efforts to shorten the incubation time and to develop assays aiming for early intraerythrocytic stages 12 , 13 . Furthermore, a method capable of detecting inhibitory action rapidly and in a stage-specific fashion might be advantageous in ex vivo field assays as well.…”

Section: Introductionmentioning

confidence: 99%

Rapid and quantitative antimalarial drug efficacy testing via the magneto-optical detection of hemozoin

Molnár

Orbán

Izrael

et al. 2020

Sci Rep

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Emergence of resistant Plasmodium species makes drug efficacy testing a crucial part of malaria control. Here we describe a novel assay for sensitive, fast and simple drug screening via the magnetooptical detection of hemozoin, a natural biomarker formed during the hemoglobin metabolism of Plasmodium species. By quantifying hemozoin production over the intraerythrocytic cycle, we reveal that hemozoin formation is already initiated by ~ 6-12 h old ring-stage parasites. We demonstrate that the new assay is capable of drug efficacy testing with incubation times as short as 6-10 h, using synchronized P. falciparum 3D7 cultures incubated with chloroquine, piperaquine and dihydroartemisinin. The determined 50% inhibitory concentrations agree well with values established by standard assays requiring significantly longer testing time. Accordingly, we conclude that magneto-optical hemozoin detection provides a practical approach for the quick assessment of drug effect with short incubation times, which may also facilitate stage-specific assessment of drug inhibitory effects.

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一种用于检测恶性疟原虫青蒿素抗性的新型表型检测方法-环状体阶段生存测定法(rsa)及其改进的综述

Cited by 11 publications

References 26 publications

Ex vivo susceptibilities of Plasmodium vivax isolates from the China-Myanmar border to antimalarial drugs and association with polymorphisms in Pvmdr1 and Pvcrt-o genes

Ex vivo susceptibilities of Plasmodium vivax isolates from the China-Myanmar border to antimalarial drugs and association with polymorphisms in Pvmdr1 and Pvcrt-o genes

In vitro susceptibility of Plasmodium falciparum isolates from the China-Myanmar border area to artemisinins and correlation with K13 mutations

Rapid and quantitative antimalarial drug efficacy testing via the magneto-optical detection of hemozoin

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