Prevalence of Placental Malaria, Plasmodium Falciparum's Histidine Rich Protein  2/3 Gene Deletions and Kelch-13 Gene Mutations among Pregnant Women in  Selected Health Facilities of Ethiopia

Banchamlak, Tegegne

dc.contributor.author	Banchamlak, Tegegne
dc.date.accessioned	2026-08-20T07:28:42Z
dc.date.available	2026-08-20T07:28:42Z
dc.date.issued	2025-11
dc.identifier.uri	http://ir.bdu.edu.et/handle/123456789/17049
dc.description.abstract	Background: Placental malaria (PM) represents a significant public health burden for both pregnant women and their fetus. The diagnosis of malaria in Ethiopia primarily relies on blood film microscopy using stained thick and thin blood films and Rapid Diagnostic Tests (RDTs). However, RDTs pfhrp2 and pfhrp3 gene deletions result in misdiagnosis of Plasmodium falciparum infection. According to WHO recommendation, artemisinin-based combination therapies (ACTs) were used as a first line anti-malarial drug in the country. On the contrary, a mutation in the PfKelch13 gene was found to be associated with artemisinin resistance in endemic countries including Ethiopia.Therefore, this study aimed to determine the prevalence of placental malaria, pfk13 gene mutation, pfhrp2/3 gene deletion and performance evaluation of lactate dehydrogenase based RDTs among symptomatic and asymptomatic pregnant women in selected health facilities (Andasa Health Center, Chis Abay Health Center, Hamusit Health Center, Kurgen Health Center, Uffa Health Center, Gambella General Hospital, Gambella Primary Hospital, and Gebretsadik Shewa Hospital) of Ethiopia. Methods: A facility based cross-sectional study was conducted from July to November 2023 at selected eight health facilities of Ethiopia. Nine hundred twenty-two pregnant women were enrolled for placental malaria prevalence study, and their placenta samples were examined for Plasmodium infection using smear microscopy and Loop-mediated isothermal amplification method. For LDH based RDT kit performance evaluation, 302 pregnant women suspected of malaria were recruited and samples were examined using blood film microscopy, CareStart RDT, Boicredit RDT and PET PCR for the detection of Plasmodium species. For hrp2/3 gene deletion and ACT drug resistance molecular marker detection, 275 pregnant women with P. falciparum infection were purposively recruited. For hrp2/3 gene deletion detection, Bio-Rad QX200 ddPCR machine was used to look at 3 targets (HRP2 exon 1, HRP 2 exon 2 and HRP 3) and their concentration in relation to tRNA ligase. Genomic DNA from P. falciparum laboratory reference strains was included in each ddPCR assay as experimental controls. The P. falciparum k13 propeller domain was amplified from genomic DNA samples by nested PCR and its products were run on 2% agarose gel and bidirectional Sanger sequencing was performed. Sociodemographic characteristics of the participants and necessary clinical data were collected using Redcap software in the anti-natal clinic and labor ward. Results: Of the total recruited pregnant women, thirteen samples (1.4%) were positive with LAMP methods of which eight (61.5%) of them had VI P. falciparum infections and five of them (38.5%) had P. vivax infections. From the total analyzed samples with ddPCR, 15% (31/207) of parasites had fully deleted hrp2/3 genes. A separate analysis of hrp2 and hrp3 gene deletion showed that 17.9% (37/207) and 41.5% (86/207) of the samples carried hrp2 and hrp3 gene deleted parasites respectively. The sensitivity and negative predictive value of BIOCREDIT Pf/Pv (pLDH/pLDH) were 55.6% (95%CI: 46.1 64.7) and 77.9% (95% CI: 74.2-81.2) against a PET-PCR reference test respectively which was higher as compared to CareStart™ Pf/Pv (HRP2/pLDH). From the total 227 sequenced samples, 18.9 % (43/227) had pfk13 gene mutations with a total of 13 SNPs (point mutations) including ten non-synonymous mutations and three synonymous mutations. The G596E, H588Y, I406R and T587I are novel non-synonymous mutations which have never been described elsewhere. Three WHO approved k13 resistance markers (R622I, A675V and R539T) were detected. Conclusion: Both P. falciparum and P. vivax, including asymptomatic infections, contribute to placental malaria. High pfhrp2/3 deletion rates and the presence of pfk13 resistance mutations pose major diagnostic and treatment concerns in Ethiopia. Continued molecular surveillance using ddPCR and whole genome sequencing is essential.	en_US
dc.language.iso	en_US	en_US
dc.subject	Biology	en_US
dc.title	Prevalence of Placental Malaria, Plasmodium Falciparum's Histidine Rich Protein 2/3 Gene Deletions and Kelch-13 Gene Mutations among Pregnant Women in Selected Health Facilities of Ethiopia	en_US
dc.type	Dissartation	en_US