Browsing by Author "Dibora Hailu"

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    Diagnostic Performance of PCR and Microscopy and Molecular Surveillance of Chloroquine Resistance-Markers in Plasmodium falciparum Clinical isolates in Hawassa Sidama Ethiopia
    (Addis Abeba University, 2026) Dibora Hailu; Samson Leta; Lemu Golassa
    Malaria is still a major public health problem in Ethiopia, particularly in areas with seasonal transmission, such as Hawassa, southern Ethiopia. Accurate diagnosis and continuous monitoring of antimalarial drug resistance are essential for effective control and elimination efforts. This study aimed to evaluate the diagnostic performance of PCR and microscopy and conduct molecular surveillance of CQ resistance markers in Plasmodium falciparum clinical isolates. A cross-sectional study was performed from September 2025 to May 2026 in selected health centers found in Hawassa, southern Ethiopia. A total of 1428 dried blood samples were collected from malaria-suspected individuals. Nested PCR targeting the 18S rRNA gene was used for species confirmation, while genotyping was conducted by restriction fragment length polymorphism (RFLP) to detect mutations in the pfcrt (K76T) and pfmdr1 (N86Y) genes. Statistical analysis was performed using R software version 4.3.2. Logistic regression was used to assess the association between infection status and participants' characteristics, and Cohen‟s kappa was used to assess agreement between microscopy and PCR. Out of 1428 samples, 546 (38.2%) were PCR positive for Plasmodium species, of which 214 were confirmed as P. falciparum. Sex was significantly associated with PCR positivity, with male participants exhibiting higher odds of infection compared to females (OR = 1.33, 95% CI: 1.04–1.69, p = 0.021). Age (as a continuous variable) showed no statistically significant association with malaria infection overall or by species (Pf and Pv), as the odds ratios were close to unity and the confidence intervals included 1 (Overall PCR: OR = 0.99, 95% CI: 0.98–1.00, P = 0.40; Pf: OR = 0.99, 95% CI: 0.98–1.00, P = 0.211; Pv: OR = 0.99, 95% CI: 0.97–1.00, P = 0.30). But age was significantly associated with microscopy positivity, (OR = 0.98, 95% CI: 0.97–0.99, p < 0.001). In conclusion, PCR showed superior diagnostic performance compared to microscopy, particularly for malaria species identification. Microscopy demonstrated moderate agreement for P. falciparum (κ = 0.426) and fair agreement for P. vivax (κ = 0.289), while overall agreement was substantial (κ = 0.611; accuracy = 82.56%). Molecular analysis revealed fixation of the pfcrt K76T mutation (100%) and absence of the pfmdr1 N86Y mutant allele, indicating persistent chloroquine resistance markers despite drug withdrawal. These findings highlight the need for continued molecular surveillance and strengthened malaria diagnosis and treatment monitoring in Ethiopia.