Development and application of ultra-sensitive tools for the detection of malaria

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dc.contributor.advisorPillai, Dylan R.
dc.contributor.authorMohon, Md Abu Naser
dc.contributor.committeememberWasmuth, James D.
dc.contributor.committeememberParkins, Michael D.
dc.date2020-02
dc.date.accessioned2020-01-15T20:39:04Z
dc.date.available2020-01-15T20:39:04Z
dc.date.issued2020-01
dc.description.abstractThe goal to eliminate malaria has been challenged by the lack of accurate diagnostic tools to identify symptomatic, asymptomatic, and drug-resistant malaria carriers. In this dissertation, we have shown the potential of the Loop-mediated Isothermal Amplification (LAMP)-based diagnostic approaches to be a powerful tool available for malaria elimination. We have validated the combination of the Non-instrumented Nucleic Acid (NINA) platform heater (PATH, Seattle) with a commercial LAMP kit (LoopAmp malaria Pan/Pf detection kit), with a view to deploying it in extremely resource-limited settings in the future. An ultrasensitive (US)-LAMP assay was also developed and validated to identify asymptomatic malaria reservoirs. Moreover, a novel strategy for detecting single nucleotide polymorphisms (SNPs) by the LAMP method was designed and deployed for spotting artemisinin resistance in P. falciparum. We conclude that the NINA-LAMP assay can be a convenient test for detecting symptomatic malaria cases with a sensitivity of 100% and specificity of 98.6% compared to the gold standard nested PCR. Additionally, the US-LAMP assay was able to achieve a limit of detection (LOD) between 25 to 100 parasites/mL from dried blood spots. We have also found that the overall prevalence of asymptomatic malaria was 22.1% in the Gambella region of Ethiopia, detected by the US-LAMP assay. The sensitivity and the specificity of the US-LAMP assay were 92.6% and 97.1%, respectively compared to an ultrasensitive quantitative reverse transcriptase PCR. Additionally, the SNP-LAMP assay was 100% sensitive and 97.3% specific to identify the C580Y mutation in the kelch 13 propeller gene, which is known as the major genetic determinant of artemisinin resistance in Southeast Asia. Furthermore, we conclude that artemisinin resistance-linked kelch 13 propeller mutations are absent in the Bangladeshi P. falciparum isolates. However, two cases of the A578S SNP in the kelch 13 propeller gene were found in those P. falciparum isolates, although this SNP was not associated with artemisinin resistance. In conclusion, as the LAMP-based diagnostic approaches are simple, low-cost, and accurate compared to currently available nucleic acid tests, they can be used at different aspects to diagnose malaria and expedite elimination.en_US
dc.identifier.citationMohon, M. A. N. (2020). Development and application of ultra-sensitive tools for the detection of malaria (Doctoral thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca.en_US
dc.identifier.doihttp://dx.doi.org/10.11575/PRISM/37455
dc.identifier.urihttp://hdl.handle.net/1880/111498
dc.language.isoengen_US
dc.publisher.facultyCumming School of Medicineen_US
dc.publisher.institutionUniversity of Calgaryen
dc.rightsUniversity of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.en_US
dc.subjectAsymptomatic malariaen_US
dc.subjectmalaria diagnosisen_US
dc.subjectultra-sensitive toolen_US
dc.subjectartemisinin resistanceen_US
dc.subjectLAMPen_US
dc.subjectSNP-LAMPen_US
dc.subjecteliminationen_US
dc.subjectpoint-of-careen_US
dc.subject.classificationMicrobiologyen_US
dc.subject.classificationBiology--Molecularen_US
dc.subject.classificationParasitologyen_US
dc.titleDevelopment and application of ultra-sensitive tools for the detection of malariaen_US
dc.typedoctoral thesisen_US
thesis.degree.disciplineMedicine – Microbiology & Infectious Diseasesen_US
thesis.degree.grantorUniversity of Calgaryen_US
thesis.degree.nameDoctor of Philosophy (PhD)en_US
ucalgary.item.requestcopytrueen_US
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