scholarly journals Analysis of nucleic acids extracted from rapid diagnostic tests reveals a significant proportion of false positive test results associated with recent malaria treatment

2021 ◽  
Author(s):  
Salome Hosch ◽  
Charlene Aya Yoboue ◽  
Olivier Tresor Donfack ◽  
Etienne A Guirou ◽  
Jean-Pierre Dangy ◽  
...  
Keyword(s):  
Quality Control ◽  
Nucleic Acids ◽  
False Positive ◽  
Diagnostic Tests ◽  
False Negative ◽  
Rapid Diagnostic Tests ◽  
Malaria Treatment ◽  
Malaria Surveillance ◽  
Bioko Island ◽  
Surveillance Programs

Surveillance programs often use malaria rapid diagnostic tests (RDTs) to determine the proportion of the population carrying parasites in their peripheral blood to assess the malaria transmission intensity. Despite an increasing number of reports on false-negative and false-positive RDT results, there is a lack of systematic quality control activities for RDTs deployed in malaria surveillance programs. Our study provides a larger scale comparative evaluation of RDTs used in the 2018 Malaria Indicator Survey (MIS) conducted on Bioko Island, Equatorial Guinea. We conducted a molecular analysis by extraction of nucleic acids from 1,800 negative and 1,065 positive RDTs followed by qPCR analysis. These results were combined with a dataset collected in a comprehensive questionnaire from each MIS participant. Of the 2,865 RDTs that were collected in 2018 on Bioko Island and analysed in our study, 4.7% had a false-negative result. These false-negative RDT results were associated with low parasite density infections. In a substantial proportion of samples, we identified masked pfhrp2 and pfhrp3 gene deletions in which at least one P. falciparum strain carried a gene deletion. Among all positive RDTs analysed, 28.4% were tested negative by qPCR and therefore considered to be false-positive. Analysing the questionnaire data collected from the participants, this high proportion of false-positive RDT results could be explained by PfHRP2 antigen persistence after recent malaria treatment. We conclude that malaria surveillance depending solely on RDTs needs well-integrated quality control procedures assessing the extend and impact of reduced sensitivity and specificity of RDTs on malaria control programs.

scholarly journals Molecular malaria surveillance using a novel protocol for extraction and analysis of nucleic acids retained on used rapid diagnostic tests

2020 ◽  
Author(s):  
Etienne A. Guirou ◽  
Tobias Schindler ◽  
Salome Hosch ◽  
Olivier Tresor Donfack ◽  
Charlene Aya Yoboue ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Diagnostic Tests ◽  
Large Scale ◽  
Rapid Diagnostic Tests ◽  
Nucleic Acid Amplification ◽  
Nucleotide Polymorphisms ◽  
Sample Collection ◽  
Malaria Surveillance ◽  
Bioko Island ◽  
Asymptomatic Patients

AbstractThe use of malaria rapid diagnostic tests (RDTs) as a source for nucleic acids that can be analyzed via nucleic acid amplification techniques has several advantages, including minimal amounts of blood, sample collection, simplified storage and shipping conditions at room temperature. We have systematically developed and extensively evaluated a procedure to extract total nucleic acids from used malaria RDTs. The co-extraction of DNA and RNA molecules from small volumes of dried blood retained on the RDTs allows detection and quantification of P. falciparum parasites from asymptomatic patients with parasite densities as low as 1 Pf/µL blood using reverse transcription quantitative PCR. Based on the extraction protocol we have developed the ENAR (Extraction of Nucleic Acids from RDTs) approach; a complete workflow for large-scale molecular malaria surveillance. Using RDTs collected during a malaria indicator survey we demonstrated that ENAR provides a powerful tool to analyze nucleic acids from thousands of RDTs in a standardized and high-throughput manner. We found several, known and new, non-synonymous single nucleotide polymorphisms in the propeller region of the kelch 13 gene among isolates circulating on Bioko Island, Equatorial Guinea.

scholarly journals Molecular malaria surveillance using a novel protocol for extraction and analysis of nucleic acids retained on used rapid diagnostic tests

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Etienne A. Guirou ◽  
Tobias Schindler ◽  
Salome Hosch ◽  
Olivier Tresor Donfack ◽  
Charlene Aya Yoboue ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Diagnostic Tests ◽  
Rapid Diagnostic Tests ◽  
Malaria Surveillance

scholarly journals PO 8271 PFHRP2 GENE DELETIONS IN PLASMODIUM FALCIPARUM AND SCHISTOSOMA MANSONI CO-INFECTIONS: AN EMERGING CHALLENGE FOR MALARIA RAPID DIAGNOSTIC TESTS

2019 ◽  
Vol 4 (Suppl 3) ◽  
pp. A25.2-A25
Author(s):  
Hilda Echelibe ◽  
Masumbe Netongo Palmer ◽  
Nji Akindeh ◽  
Wilfred Mbacham
Keyword(s):  
Plasmodium Falciparum ◽  
Schistosoma Mansoni ◽  
Diagnostic Tests ◽  
False Negative ◽  
Rapid Diagnostic Tests ◽  
Sub Saharan Africa ◽  
P Value ◽  
Gene Deletions ◽  
Malaria Rdts ◽  
Malaria Rapid Diagnostic Tests

BackgroundMalaria and schistosomiasis are infections that have a great impact in sub-Saharan Africa based on their high morbidity and mortality rates. We suggest the possibility that the microenvironment created from interactions between the parasites involved generates a pressure on the malaria parasite which could in turn favour the parasite’s adaptation or escape through Pfhrp2 gene deletions. Thus, this study aimed at determining the association between the co-infection with both parasites and false-negative PfHRP2-based malaria rapid diagnostic tests which occur because of these deletions.MethodsThis pilot study was conducted in a total of 149 children aged 7–17 years living in Yorro, located in the Mbam-Inoubou division of the Center region of Cameroon. We collected fresh stool samples from each participant to identify Schistosoma mansoni (Sm) eggs by Kato Katz method and blood samples to identify the ring stages of Plasmodium falciparum (Pf) by thick smear. Malaria rapid diagnostic test and Pfhrp2 gene polymerase chain reaction were performed. The association between the co-infection with Sm/Pf and the false-negative malaria RDTs was determined by the Fisher’s exact test. A p value<0.05 was considered statistically significant.ResultsOur results showed that samples were singly infected with Sm, Pf, co-infected (Sm/Pf) and negative for both infections at frequencies of 12%, 43%, 30.2% and 14.8% respectively. False-negative PfHRP2-based RDTs were observed in 4.7% of the participants. A higher frequency (5/7) of the cases with false-negative malaria RDTs were co-infected with Sm/Pf. A p value of 0.027 showed statistical significance in the association of Sm/Pf co-infection and false-negative PfHRP2-based RDTs.ConclusionA significant association of Plasmodium falciparum and Schistosoma mansoni co-infection with false-negative PfHRP2-based RDTs supports the case for a plausible implication of Pfhrp2 gene deletions, with consequences for malaria rapid diagnostic testing.

scholarly journals An assessment of false positive rates for malaria rapid diagnostic tests caused by non-Plasmodium infectious agents and immunological factors

PLoS ONE ◽  
2018 ◽  
Vol 13 (5) ◽  
pp. e0197395 ◽  
Author(s):  
Michelle L. Gatton ◽  
Sadmir Ciketic ◽  
John W. Barnwell ◽  
Qin Cheng ◽  
Peter L. Chiodini ◽  
...  
Keyword(s):  
False Positive ◽  
Diagnostic Tests ◽  
Rapid Diagnostic Tests ◽  
Infectious Agents ◽  
Malaria Rapid Diagnostic Tests ◽  
Immunological Factors

scholarly journals Prozone-like phenomenon in travellers with fatal malaria: report of two cases

2015 ◽  
Vol 9 (03) ◽  
pp. 321-324 ◽  
Author(s):  
Lurdes Santos ◽  
Nuno Rocha Pereira ◽  
Paulo Andrade ◽  
Paulo Figueiredo Dias ◽  
Carlos Lima Alves ◽  
...  
Keyword(s):  
Diagnostic Tests ◽  
False Negative ◽  
Specific Antigen ◽  
Malaria Diagnosis ◽  
High Sensitivity ◽  
Rapid Diagnostic Tests ◽  
Microscopy Observation ◽  
Negative Results ◽  
False Negative Results ◽  
Potential Benefits

Malaria diagnosis remains a concern in non-endemic countries, with rapid diagnosis being crucial to improve patients’ outcome. Rapid diagnostic tests have high sensitivity but they also have flaws and false-negative results that might jeopardize malaria diagnosis. Some false-negative results might relate to a prozone-like effect. The authors describe two patients with false-negative rapid diagnostic tests in which a prozone-like effect might have been involved. The authors highlight that these tests should not be used without accompanying light microscopy observation of blood films and discuss potential benefits of using rapid diagnostic tests with more than one specific antigen for Plasmodium falciparum.

scholarly journals Health workers’ compliance to rapid diagnostic tests (RDTs) to guide malaria treatment: a systematic review and meta-analysis

2016 ◽  
Vol 15 (1) ◽  
Author(s):  
Alinune N. Kabaghe ◽  
Benjamin J. Visser ◽  
Rene Spijker ◽  
Kamija S. Phiri ◽  
Martin P. Grobusch ◽  
...  
Keyword(s):  
Systematic Review ◽  
Diagnostic Tests ◽  
Health Workers ◽  
Meta Analysis ◽  
Rapid Diagnostic Tests ◽  
Malaria Treatment

scholarly journals Plasmodium falciparum Allelic Diversity: A Comparison of DNA Extraction from Isolates Collected on Rapid Diagnostic Tests (Rdts) and Filter Paper

2021 ◽  
Author(s):  
Denise Patricia Mawili-Mboumba ◽  
Marie Louise Tshibola Mbuyi ◽  
Noe Patrick M’bondoukwe ◽  
Marielle Karine Bouyou-Akotet
Keyword(s):  
Plasmodium Falciparum ◽  
Nucleic Acids ◽  
Filter Paper ◽  
Diagnostic Tests ◽  
Allelic Diversity ◽  
Storage Conditions ◽  
Rapid Diagnostic Tests ◽  
Sample Collection ◽  
Filter Papers

Background: To perform molecular epidemiologic studies based on large cohorts, material such as RDTs or filter papers are useful for biological sample collection and extraction of RNA or DNA of good quality. Thus, we aimed to assess the quality of DNA extracted from malaria rapid diagnostic tests (RDTs) stored at various temperatures for the analysis of Plasmodium falciparum genetic diversity. Methods: Febrile patients benefitted from free malaria diagnosis using microscopy in a malaria sentinel site, at the Regional Hospital Estuaire-Melen, in Gabon, in 2015. P. falciparum isolates were collected onto one filter paper and 2 similar RDTs devices (Acon®) per patient. Nucleic acids were extracted with QiAmp Qiagen kit from paper and RDTs and the quality of the DNA was analyzed by msp1 gene amplification. Results: Msp1gene amplification was achieved in nucleic acids extracted from all filter papers and RDTs devices (n = 45, 100%). K1 alleles were detected in 93.3% (n = 42/45) of the samples and Mad20 alleles in 73.3% (n = 33/45). The number and the intensity of K1 and/or Mad20 fragments were comparable according to the sample collection material and the storage conditions (room temperature vs -20°C) of the samples. The size of the fragments indicating allelic diversity was comparable in 80% (n=36) of the samples. Conclusion: These data show that RDTs are a valuable source of DNA for malaria parasite genetic polymorphism analysis. Storage conditions of the devices did not influence the quality of DNA extracted from RDTs device, although some alleles may be missed.

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