Distinction of Plasmodium ovale wallikeri and Plasmodium ovale curtisi using quantitative Polymerase Chain Reaction with High Resolution Melting revelation

Joste, Valentin; Kamaliddin, Claire; Kendjo, Éric; Hubert, Véronique; Argy, Nicolas; Houzé, Sandrine

doi:10.1038/s41598-017-18026-1

Cited by 23 publications

(13 citation statements)

References 30 publications

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“…We present here the results of the implementation of a PCR-HRM assay that allows the rapid and accurate detection and identification of three Plasmodium spp., in either clinical samples or Anopheles vector samples. Our assay differs from those PCR-HRM already published 35,36,42 that have only been used to detect Plasmodium spp. in clinical samples.…”

Section: Discussionmentioning

confidence: 87%

A new high-resolution melting analysis for the detection and identification of Plasmodium in human and Anopheles vectors of malaria

Murillo

Muskus

Agudelo

et al. 2019

Sci Rep

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Among vector-borne diseases malaria is the leading cause of morbidity in the world, with more than 200 million cases per year and a large number of deaths. The techniques traditionally used for the detection of Plasmodium in humans and Anopheles mosquitoes include microscopy, IRMA, ELISA, antibody or molecular assays, and anopheline dissection. However, these techniques are limited by their requirement of skilled personnel, low sensitivity or long processing times. A PCR-based high-resolution melting (PCR-HRM) analysis was developed for the detection and identification of P. falciparum, P. vivax and P. malariae that infect humans and Anopheles. In 41 human samples PCR-HRM detected 14 samples positive for P. vivax, 17 for P. falciparum, three for P. malariae, three mixed infections for P. vivax/P. malariae and four negative samples. Whereas benchmarking assays of microscopy and nested PCR had false positive detections. Additionally, PCR-HRM was able to detect natural infection with Plasmodium spp. in An. darlingi and An. mattogrossensis. The PCR-HRM presented is the first single assay developed for the detection and identification of P. vivax, P. falciparum and/or P. malariae in human and Anopheles. This method improves on currently available assays as it is easy-to-use, rapid, sensitive and specific with a low risk of contamination.

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Section: Discussionmentioning

confidence: 87%

A new high-resolution melting analysis for the detection and identification of Plasmodium in human and Anopheles vectors of malaria

Murillo

Muskus

Agudelo

et al. 2019

Sci Rep

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“…High-resolution melting (HRM) detection based on qPCR was developed in our laboratory (17) and was performed to quantify P. falciparum DNA in samples using a calibration range obtained from serial dilution of DNA in free water (equivalent of 40,000, 4,000, 400, 40, 20, 8, and 4 parasites/l). The amplified sequence melting temperature (T m ) was recorded to ensure reaction specificity.…”

Section: Rapid Diagnosis Testmentioning

confidence: 99%

Evaluation of PCR To Monitor Plasmodium falciparum Treatment Efficacy in a Nonendemicity Setting

et al. 2019

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Adequate clinical and parasitological response (ACPR) after malaria treatment remains challenging to assess in settings of malaria nonendemicity. Biological evaluation of parasitological clearance relies on microscopic investigation of thick blood smears, which is a specific technique that not all diagnosis laboratories are able to perform. Rapid diagnosis tests (RDTs) and molecular biology techniques are proposed as alternatives to microscope conventional techniques; however, their performance for treatment efficacy evaluation is controversial. We present here a retrospective comparative study for RDT and PCR (nested and high-resolution-melting quantitative PCR [HRM-qPCR]) evaluation of ACPR in a nonendemicity context. Blood samples from 133 patients presenting a Plasmodium falciparum monoinfection were included. Samples obtained at the time of diagnosis and at 3, 7, and 28 days after diagnosis were investigated. Histidine-rich protein 2 (HRP-2)-based RDT results remained positive in 51% of cases 28 days after diagnosis and appropriate therapeutic management. Parasite DNA was detected by the two PCR techniques (nested PCR and HRM-qPCR) in 12% and 10% of samples 28 days after treatment initiation, respectively. No therapeutic failure was recorded in the studied patients. Persistence of positive signal might reflect the presence of circulating asexual parasites or persistence of HRP-2 and parasitic DNA in patient’s peripheral blood after parasitic clearance.

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“…Various PCR approaches for the detection and discrimination of P. o. curtisi and P. o. wallikeri have been described [18, 32–36], including TaqMan qPCR [18, 32–34], semi-nested PCR [37], and nested PCR [38] as well as quantitative and high-resolution melting approaches with detection limits as low as 1 parasite/µL [39]. Multilocus genotyping has also been applied for P. o. curtisi and P. o. wallikeri discrimination [23, 38].…”

Section: Introductionmentioning

confidence: 99%

A comparison of two PCR protocols for the differentiation of Plasmodium ovale species and implications for clinical management in travellers returning to Germany: a 10-year cross-sectional study

Frickmann

Wegner

Ruben

et al. 2019

Malar J

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Background: To assess the occurrence of Plasmodium ovale wallikeri and Plasmodium ovale curtisi species in travellers returning to Germany, two real-time PCR protocols for the detection and differentiation of the two P. ovale species were compared. Results of parasite differentiation were correlated with patient data. Methods: Residual nucleic acid extractions from EDTA blood samples of patients with P. ovale spp. malaria, collected between 2010 and 2019 at the National Reference Centre for Tropical Pathogens in Germany, were subjected to further parasite discrimination in a retrospective assessment. All samples had been analysed by microscopy and by P. ovale spp.-specific real-time PCR without discrimination on species level. Two different real-time PCR protocols for species discrimination of P. o. curtisi and P. o. wallikeri were carried out. Results were correlated with patient data on gender, age, travel destination, thrombocyte count, and duration of parasite latency. Results: Samples from 77 P. ovale spp. malaria patients were assessed, with a male:female ratio of about 2:1 and a median age of 30 years. Parasitaemia was low, ranging from few visible parasites up to 1% infected erythrocytes. Discriminative real-time PCRs revealed 41 cases of P. o. curtisi and 36 cases of P. o. wallikeri infections. Concordance of results by the two PCR approaches was 100%. Assessment of travel destinations confirmed coexistence of P. o. curtisi and P. o. wallikeri over a wide range of countries in sub-Saharan Africa. Latency periods for the two P. ovale species were similar, with median values of 56.0 days for P. o. curtisi and 58.0 days for P. o. wallikeri; likewise, there was no statistically significant difference in thrombocyte count with median values of 138.5/µL for patients with P. o. curtisi and 152.0/µL for P. o. wallikeri-infected patients. Conclusions: Two different real-time PCR protocols were found to be suitable for the discrimination of P. o. curtisi and P. o. wallikeri with only minor differences in sensitivity. Due to the overall low parasitaemia and the lack of differences in severity-related aspects like parasite latency periods or thrombocyte counts, this study supports the use of P. ovale spp. PCR without discrimination on species level to confirm the diagnosis and to inform clinical management of malaria in these patients.

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Distinction of Plasmodium ovale wallikeri and Plasmodium ovale curtisi using quantitative Polymerase Chain Reaction with High Resolution Melting revelation

Cited by 23 publications

References 30 publications

A new high-resolution melting analysis for the detection and identification of Plasmodium in human and Anopheles vectors of malaria

A new high-resolution melting analysis for the detection and identification of Plasmodium in human and Anopheles vectors of malaria

Evaluation of PCR To Monitor Plasmodium falciparum Treatment Efficacy in a Nonendemicity Setting

A comparison of two PCR protocols for the differentiation of Plasmodium ovale species and implications for clinical management in travellers returning to Germany: a 10-year cross-sectional study

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