Comparison of three diagnostic methods (microscopy, RDT, and PCR) for the detection of malaria parasites in representative samples from Equatorial Guinea

Berzosa, Pedro; Lucio, Aida de; Romay‐Barja, María; Herrador, Zaida; González, Verónica; García, Luz; Fernández-Martínez, Amalia; Santana-Morales, Maria A.; Ncogo, Policarpo; Valladares, Basilio; Riloha, Matilde; Benito, Agustín

doi:10.1186/s12936-018-2481-4

Cited by 229 publications

(260 citation statements)

References 37 publications

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“…Therefore, our study corroborates the limitations of microscopy in the detection of P. vivax and P. knowlesi, that have been clearly demonstrated in large scale studies in S. E. Asia [13,14] Our study also suggests that P. knowlesi, which is thought to be predominantly a S. E. Asian parasite, is now also emerging in the Indian mainland.…”

Section: Discussionsupporting

confidence: 88%

“…Asia [8][9][10][11] with a single report from the Andaman and Nicobar Islands of India [12]. In many malaria cases in endemic areas severe malaria symptoms such as AKI manifest, but malaria diagnosis by microscopy or RDT is made only in a proportion of these cases, and the rest remain as occult malaria infections [13][14][15][16]. An earlier study from our center, based on which this study has been built, had reported the presence of peripheral P. vivax infection in patients with AKI [17].…”

Section: Introductionmentioning

confidence: 99%

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Renal detection of Plasmodium falciparum, Plasmodium vivax and Plasmodium knowlesi in malaria associated acute kidney injury: a retrospective case–control study

et al. 2020

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Objective: Acute kidney injury (AKI) is a frequent presentation in malaria infections. Several cases of AKI that are accompanied by clinical symptoms of malaria infection, such as fever, nausea, respiratory distress, and anemia remain undiagnosed due to challenges in accurate diagnosis using peripheral blood microscopy and rapid diagnostic tests that are currently used in clinical settings. This is particularly true for P. vivax and P. knowlesi infections. As a result, these patients are not able to receive anti-malarial therapy in a timely manner. The objective of the present study was to investigate if patients presenting with AKI harbored any of the five human Plasmodium species (P. falciparum, P. vivax, P. knowlesi, P. malariae, and P. ovale) within their renal tissues. Results: We found that renal biopsies from malaria associated AKI patients harbor the human malaria parasites P. falciparum, P. vivax and P. knowlesi as mono-and mixed species infections. Presence of microvascular injury in a majority of the malaria associated AKI cases suggested vascular involvement of P. vivax and P. knowlesi. This research note also highlights P. knowlesi as an emerging pathogen in the Indian subcontinent.

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Section: Discussionsupporting

confidence: 88%

Section: Introductionmentioning

confidence: 99%

Renal detection of Plasmodium falciparum, Plasmodium vivax and Plasmodium knowlesi in malaria associated acute kidney injury: a retrospective case–control study

et al. 2020

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“…DNA-based malaria assays in general, and PCR in particular, are significantly more sensitive than microscopy and RDTs. Limit of detection (LOD) for PCR is typically 1-5 parasites/µL [13][14][15][16][17] compared to 50-500 parasites/µL for microscopy [13,14,18] and more than 100 parasites/µL for RDT's [14,19]. Depending on the assay, PCR can be used for accurate species identification, detection of mixed infections, and parasite density estimation [20].…”

Section: Introductionmentioning

confidence: 99%

Use of real-time multiplex PCR, malaria rapid diagnostic test and microscopy to investigate the prevalence of Plasmodium species among febrile hospital patients in Sierra Leone

Łęski

Taitt

Swaray

et al. 2020

Malar J

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Background: Malaria continues to affect over 200 million individuals every year, especially children in Africa. Rapid and sensitive detection and identification of Plasmodium parasites is crucial for treating patients and monitoring of control efforts. Compared to traditional diagnostic methods such as microscopy and rapid diagnostic tests (RDTs), DNA based methods, such as polymerase chain reaction (PCR) offer significantly higher sensitivity, definitive discrimination of Plasmodium species, and detection of mixed infections. While PCR is not currently optimized for routine diagnostics, its role in epidemiological studies is increasing as the world moves closer toward regional and eventually global malaria elimination. This study demonstrates the field use of a novel, ambient temperature-stabilized, multiplexed PCR assay in a small hospital setting in Sierra Leone.Methods: Blood samples from 534 febrile individuals reporting to a hospital in Bo, Sierra Leone, were tested using three methods: a commercial RDT, microscopy, and a Multiplex Malaria Sample Ready (MMSR) PCR designed to detect a universal malaria marker and species-specific markers for Plasmodium falciparum and Plasmodium vivax. A separate PCR assay was used to identify species of Plasmodium in samples in which MMSR detected malaria, but was unable to identify the species.Results: MMSR detected the presence of any malaria marker in 50.2% of all tested samples with P. falciparum identified in 48.7% of the samples. Plasmodium vivax was not detected. Testing of MMSR P. falciparum-negative/universal malaria-positive specimens with a panel of species-specific PCRs revealed the presence of Plasmodium malariae (n = 2) and Plasmodium ovale (n = 2). The commercial RDT detected P. falciparum in 24.6% of all samples while microscopy was able to detect malaria in 12.8% of tested specimens.

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“…A total of 1,724 blood samples were diagnosed by microscopy, SnM-PCR and RDT [21]. The Plasmodium sp.…”

Section: Resultsmentioning

confidence: 99%

“…The samples were collected from a cross-sectional survey conducted in June-August 2013 in Bata as part of a project called "PREVAMAL". A total of 1,741 individuals (1,043 in urban settings and 698 in rural) were recruited [20,21]. Figure 1 shows the different locations where the samples were collected.…”

Section: Study Populationmentioning

confidence: 99%

First evidence of the deletion in the Pfhrp2 and Pfhrp3 genes in P. falciparum from Equatorial Guinea

Berzosa

González

Taravillo

et al. 2019

Preprint

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Background: WHO recommends RDTs as a good alternative malaria-diagnosis method in remote parts of sub-Saharan Africa. The majority of commercial RDTs currently available detect the P. falciparum protein histidine-rich protein 2 (PfHRP2). There have also been recent reports of Pfhrp2 deletions being found in parasites collected from several African countries. WHO has concluded that the lacking the Pfhrp2 gene must be monitored in Africa. The purpose of the study was to analyze why the samples that were positive by PCR were negative by RDTs; and, therefore, to determine whether there have been deletions in the Pfhrp2 and/or Pfhrp3 genes. Methods: Malaria NM-PCR was carried out to all the samples collected in the field. A group of 128 samples was positive by PCR but negatives by RDT, these samples were classified as RDT false-negatives. It was carried out a PCR for exon2 of Pfhrp2 and Pfhrp3 genes to detect the presence or absence of these two genes. Frequencies with 95% confidence intervals (CIs) were used for categorical variables. Associations were assessed by the chi-square test or Fisher´s exact test. The level of significance was set at p ≤ 0.05. Statistical analyses were performed using the software package SPSSv.15.0. Sensitivity and specificity calculations were performed using Epidat 3.1 software. Results: After the PCR, 81 samples were identified (4.7%, 95%CI: 3.8-5.8) which had deletion in both genes, Pfhrp2 and Pfhrp3. Overall, however, 11 samples (0.6%, 95%CI: 0.36-1.14) had deletion only in Pfhrp2 but not in Pfhrp3, and 15 (0.9%, 95%CI: 0.6-1.5) presented with deletion only in Pfhrp3 but not in Pfhrp2. Considering the Pfhrp2 gene separately, within the total of 1,724 samples, 92 (5.3%, 95%CI: 4.37-6.5) had evidence of deletion. Conclusion: The present study provides the first evidence of deletion in the Pfhrp2 and Pfhrp3 genes in P. falciparum isolates from Equatorial Guinea. However, larger studies across different regions within the country and across different seasonal profiles are needed to determine the full extent of Pfhrp2- and Pfhrp3-deletion. it would be strongly recommendable to implement an active surveillance program in order to detect any increases in Pfhrp2- and Pfhrp3-deletion frequencies.

show abstract

Comparison of three diagnostic methods (microscopy, RDT, and PCR) for the detection of malaria parasites in representative samples from Equatorial Guinea

Cited by 229 publications

References 37 publications

Renal detection of Plasmodium falciparum, Plasmodium vivax and Plasmodium knowlesi in malaria associated acute kidney injury: a retrospective case–control study

Renal detection of Plasmodium falciparum, Plasmodium vivax and Plasmodium knowlesi in malaria associated acute kidney injury: a retrospective case–control study

Use of real-time multiplex PCR, malaria rapid diagnostic test and microscopy to investigate the prevalence of Plasmodium species among febrile hospital patients in Sierra Leone

First evidence of the deletion in the Pfhrp2 and Pfhrp3 genes in P. falciparum from Equatorial Guinea

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