Molecular Diagnosis of Malaria by Photo-Induced Electron Transfer Fluorogenic Primers: PET-PCR

Lucchi, Naomi W.; Jothikumar, Narayanan; Karell, Mara A.; Xayavong, Maniphet; Kariuki, Simon; Dasilva, Alexandre J; Hill, Vincent R.; Udhayakumar, Venkatachalam

doi:10.1371/journal.pone.0056677

Cited by 118 publications

(139 citation statements)

References 22 publications

(28 reference statements)

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“…Genomic DNA was isolated from blood spots taken at enrollment (day 0) using a QIAamp DNA minikit (Qiagen, Valencia, CA). Samples were screened using the multiplex photo-induced electron transfer (PET)-PCR (15). For each sample, duplicate PET-PCRs were run with 5 l of DNA template used in the PCR.…”

Section: Methodsmentioning

confidence: 99%

Molecular Profile of Malaria Drug Resistance Markers of Plasmodium falciparum in Suriname

Chenet

Okoth

Kelley

et al. 2017

Antimicrob Agents Chemother

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In Suriname, an artesunate monotherapy therapeutic efficacy trial was recently conducted to evaluate partial artemisinin resistance emerging in Plasmodium falciparum. We genotyped the PfK13 propeller domain of P. falciparum in 40 samples as well as other mutations proposed to be associated with artemisininresistant mutants. We did not find any mutations previously associated with artemisinin resistance in Southeast Asia, but we found fixed resistance mutations for chloroquine (CQ) and sulfadoxine-pyrimethamine. Additionally, the PfCRT C350R mutation, associated with reversal of CQ resistance and piperaquine-selective pressure, was present in 62% of the samples. Our results from neutral microsatellite data also confirmed a high parasite gene flow in the Guiana Shield. Although recruiting participants for therapeutic efficacy studies is challenging in areas where malaria endemicity is very low due to the low number of malaria cases reported, conducting these studies along with molecular surveillance remains essential for the monitoring of artemisinin-resistant alleles and for the characterization of the population structure of P. falciparum in areas targeted for malaria elimination.

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Section: Methodsmentioning

confidence: 99%

Molecular Profile of Malaria Drug Resistance Markers of Plasmodium falciparum in Suriname

Chenet

Okoth

Kelley

et al. 2017

Antimicrob Agents Chemother

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“…Genomic DNA was isolated from dried blood spots using the QIAamp DNA blood minikit (Qiagen, Valencia, CA) per the manufacturer's instruction. Confirmation of P. falciparum infection and DNA quality assessment were conducted using photoinduced electron transfer (PET)-PCR (15).…”

Section: Methodsmentioning

confidence: 99%

Efficacy of Artemether-Lumefantrine and Dihydroartemisinin-Piperaquine for Treatment of Uncomplicated Malaria in Children in Zaire and Uíge Provinces, Angola

Pluciński

Talundzic

Morton

et al. 2015

Antimicrob Agents Chemother

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The development of resistance to antimalarials is a major challenge for global malaria control. Artemisinin-based combination therapies, the newest class of antimalarials, are used worldwide but there have been reports of artemisinin resistance in Southeast Asia. In February through May 2013, we conducted open-label, nonrandomized therapeutic efficacy studies of artemetherlumefantrine (AL) and dihydroartemisinin-piperaquine (DP) in Zaire and Uíge Provinces in northern Angola. The parasitological and clinical responses to treatment in children with uncomplicated Plasmodium falciparum monoinfection were measured over 28 days, and the main outcome was a PCR-corrected adequate clinical and parasitological response (ACPR) proportion on day 28. Parasites from treatment failures were analyzed for the presence of putative molecular markers of resistance to lumefantrine and artemisinins, including the recently identified mutations in the K13 propeller gene. In the 320 children finishing the study, 25 treatment failures were observed: 24 in the AL arms and 1 in the DP arm. The PCR-corrected ACPR proportions on day 28 for AL were 88% (95% confidence interval [CI], 78 to 95%) in Zaire and 97% (91 to 100%) in Uíge. For DP, the proportions were 100% (95 to 100%) in Zaire, and 100% (96 to 100%) in Uíge. None of the treatment failures had molecular evidence of artemisinin resistance. In contrast, 91% of AL late-treatment failures had markers associated with lumefantrine resistance on the day of failure. The absence of molecular markers for artemisinin resistance and the observed efficacies of both drug combinations suggest no evidence of artemisinin resistance in northern Angola. There is evidence of increased lumefantrine resistance in Zaire, which should continue to be monitored.

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“…Many of these molecular tools for malaria diagnosis range from conventional PCR-based assays, real-time PCR assay, to isothermal assays [10]. …”

Section: Introductionmentioning

confidence: 99%

Evaluation of the utility value of three diagnostic methods in the detection of malaria parasites in endemic area

Ugah¹,

Alo²,

Owolabi³

et al. 2017

Malar J

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BackgroundMalaria is a debilitating disease with high morbidity and mortality in Africa, commonly caused by different species of the genus Plasmodium in humans. Misdiagnosis is a major challenge in endemic areas because of other disease complications and technical expertise of the medical laboratory staff. Microscopic method using Giemsa-stained blood film has been the mainstay of diagnosis of malaria. However, since 1993 when rapid diagnostic test (RDT) kits were introduced, they have proved to be effective in the diagnosis of malaria. This study was aimed at comparing the accuracy of microscopy and RDTs in the diagnosis of malaria using nested PCR as the reference standard. Four hundred and twenty (420) venous blood specimens were collected from patients attending different General Hospitals in Ebonyi State with clinical symptoms of malaria. The samples were tested with Giemsa-stained microscopy and three RDTs. Fifty specimens were randomly selected for molecular analysis.ResultsUsing different diagnostic methods, the prevalence of malaria among the subjects studied was 25.95% as detected by microscopy, prevalence found among the RDTs was 22.90, 15.20 and 54.80% for Carestart, SD Bioline PF and SD Bioline PF/PV, respectively. Molecular assay yielded a prevalence of 32%. The major specie identified was Plasmodium falciparum; there was co-infection of P. falciparum with Plasmodium malariae and Plasmodium ovale. The sensitivity and specificity of microscopy was 50.00 and 70.59% while that of the RDTs were (25.00 and 85.29%), (25.00 and 94.12%) and (68.75 and 52.94%) for Carestart, SD Bioline PF and SD Bioline PF/PV, respectively. Cohen’s kappa coefficient was used to measure the level of agreement of the methods with nested PCR. Microscopy showed a moderate measure of agreement (k = 0.491), Carestart showed a good measure of agreement (k = 0.611), SD Bioline PF showed a fair measure of agreement (k = 0.226) while SD Bioline PF/PV showed a poor measure of agreement (k = 0.172).ConclusionsThis study recommends that the policy of malaria diagnosis be changed such that the routine diagnosis of malaria is done by a combination of both microscopy and a RDT kit of high sensitivity and specificity so as to complement the errors associated with either of the methods. The finding of P. ovale in the study area necessitates an expanded molecular epidemiology of malaria within the study area.

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Molecular Diagnosis of Malaria by Photo-Induced Electron Transfer Fluorogenic Primers: PET-PCR

Cited by 118 publications

References 22 publications

Molecular Profile of Malaria Drug Resistance Markers of Plasmodium falciparum in Suriname

Molecular Profile of Malaria Drug Resistance Markers of Plasmodium falciparum in Suriname

Efficacy of Artemether-Lumefantrine and Dihydroartemisinin-Piperaquine for Treatment of Uncomplicated Malaria in Children in Zaire and Uíge Provinces, Angola

Evaluation of the utility value of three diagnostic methods in the detection of malaria parasites in endemic area

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