Novel Plasmodium falciparum k13 gene polymorphisms from Kisii County, Kenya during an era of artemisinin-based combination therapy deployment.

BACKGROUND: Currently, chemotherapy stands out as the major malaria intervention strategy, however, anti-malarial resistance may hamper global elimination programs. Artemisinin-based combination therapy (ACT) stands as the drug of choice for the treatment of Plasmodium falciparum malaria. Plasmodium falciparum kelch13 gene mutations are associated with artemisinin resistance. Thus, this study was aimed at evaluating the circulation of P. falciparum k13 gene polymorphisms from Kisii County, Kenya during an era of ACT deployment. METHODS: Participants suspected to have malaria were recruited. Plasmodium falciparum was confirmed using the microscopy method. Malaria-positive patients were treated with artemether-lumefantrine (AL). Blood from participants who tested positive for parasites after day 3 was kept on filter papers. DNA was extracted using chelex-suspension method. A nested polymerase chain reaction (PCR) was conducted and the second-round products were sequenced using the Sanger method. Sequenced products were analysed using DNAsp 5.10.01 software and then blasted on the NCBI for k13 propeller gene sequence identity using the Basic Local Alignment Search Tool (BLAST). To assess the selection pressure in P. falciparum parasite population, Tajima' D statistic and Fu & Li's D test in DnaSP software 5.10.01 was used. RESULTS: Out of 275 enrolled participants, 231 completed the follow-up schedule. 13 (5.6%) had parasites on day 28 hence characterized for recrudescence. Out of the 13 samples suspected of recrudescence, 5 (38%) samples were positively amplified as P. falciparum, with polymorphisms in the k13-propeller gene detected. Polymorphisms detected in this study includes R539T, N458T, R561H, N431S and A671V, respectively. The sequences have been deposited in NCBI with bio-project number PRJNA885380 and accession numbers SAMN31087434, SAMN31087433, SAMN31087432, SAMN31087431 and SAMN31087430 respectively. CONCLUSIONS: WHO validated polymorphisms in the k13-propeller gene previously reported to be associated with ACT resistance were not detected in the P. falciparum isolates from Kisii County, Kenya. However, some previously reported un-validated k13 resistant single nucleotide polymorphisms were reported in this study but with limited occurrences. The study has also reported new SNPs. More studies need to be carried out in the entire country to understand the association of reported mutations if any, with ACT resistance.

Trend of Malaria Burden Among Residents of Kisii County, Kenya After More Than a Decade Usage of Artemisinin Combined Therapies, 11-Year Laboratory Based Retrospective Study.

Background: Malaria remains a major vector borne disease globally, with the majority of the casualties reported in Africa. Despite this fact, there is drastic reduction in malaria infection using Artemisinin combined therapies (ACTs). Malaria is characterized by significant inconsistency in different geographical locations due to different confounding factors. There is need to identify zone-specific malaria trends and interventions to completely eliminate the disease. Thus the study was aimed at assessing the 11-year trend of microscopically confirmed malaria cases in Kisii County, Kenya, so as to devise area-specific evidence-based interventions, informed decisions, and to track the effectiveness of malaria control programs. Methods: This was a retrospective study carried out to determine 11-year malaria trend rates centered on the admission and laboratory records from health facilities located at four Sub-Counties in Kisii County, Kenya. Parasitological positivity rates of malaria were determined by comparing with the register records in health facilities which recorded confirmed malaria cases with the total number of monthly admissions over the entire year. Data was analyzed by using descriptive tools and chi-square test. Results: There were 36,946 suspect cases, with 8449 (22.8%) confirmed malaria cases reported in this study. The overall malaria slide positivity rate over the last 11 years in the study area was 22.6%. The months of April and August showed the largest number of malaria cases (63%). The age group of ≥18 years contained the most positive confirmed cases, having a prevalence rate of 2953 (35.45%). Out of the confirmed malaria cases, 2379 (28.1%) were males and 6070 (71.9%) were females The highest malaria prevalence rate was recorded in 2014, with Marani Sub-County recording the highest positivity rate of 37.94%. Conclusion: From the observed trends, malaria prevalence and transmission still remains stable in the study area. Thus more interventions need to be scaled up.