In Uganda, Artemether-Lumefantrine and Artesunate are recommended for uncomplicated and severe malaria respectively, but are currently threatened by parasite resistance. Genetic and epigenetic factors play a role in predisposing Plasmodium falciparum parasites to acquiring Pfkelch13 (K13) mutations associated with delayed artemisinin parasite clearance as reported in Southeast Asia. In this study, we report on the prevalence of mutations in the K13, pfmdr-2 (P. falciparum multidrug resistance protein 2), fd (ferredoxin), pfcrt (P. falciparum chloroquine resistance transporter), and arps10 (apicoplast ribosomal protein S10) genes in Plasmodium falciparum parasites prior to (2005) and after (2013) introduction of artemisinin combination therapies for malaria treatment in Uganda. A total of 200 P. falciparum parasite DNA samples were screened. Parasite DNA was extracted using QIAamp DNA mini kit (Qiagen, GmbH, Germany) procedure. The PCR products were sequenced using Sanger dideoxy sequencing method. Of the 200 P. falciparum DNA samples screened, sequencing for mutations in K13, pfmdr-2, fd, pfcrt, arps10 genes was successful in 142, 186, 141, 128 and 74 samples respectively. Overall, we detected six (4.2%, 6/142; 95%CI: 1.4–7.0) K13 single nucleotide polymorphisms (SNPs), of which 3.9% (2/51), 4.4% (4/91) occurred in 2005 and 2013 samples respectively. All four K13 SNPs in 2013 samples were non-synonymous (A578S, E596V, S600C and E643K) while of the two SNPs in 2005 samples, one (Y588N) is non-synonymous and the other (I587I) is synonymous. There was no statistically significant difference in the prevalence of K13 (p = 0.112) SNPs in the samples collected in 2005 and 2013. The overall prevalence of SNPs in pfmdr-2 gene was 39.8% (74/186, 95%CI: 25.1–50.4). Of this, 4.2% (4/95), 76.9% (70/91) occurred in 2005 and 2013 samples respectively. In 2005 samples only one SNP, Y423F (4.2%, 4/95) was found while in 2013, Y423F (38.5%, 35/91) and I492V (38.5%, 35/91) SNPs in the pfmdr-2 gene were found. There was a statistically significant difference in the prevalence of pfmdr-2 SNPs in the samples collected in 2005 and 2013 (p<0.001). The overall prevalence of arps10 mutations was 2.7% (2/72, 95%CI: 0.3–4.2). Two mutations, V127M (4.5%: 1/22) and D128H (4.5%: 1/22) in the arps10 gene were each found in P. falciparum parasite samples collected in 2013. There was no statistically significant difference in the prevalence of arps10 SNPs in the samples collected in 2005 and 2013 (p = 0.238). There were more pfmdr-2 SNPs in P. falciparum parasites collected after introduction of Artemisinin combination therapies in malaria treatment. This is an indicator of the need for continuous surveillance to monitor emergence of molecular markers of artemisinin resistance and its potential drivers in malaria affected regions globally.

Background Substandard anti-malarial agents pose a significant challenge to effective malaria control and elimination efforts especially in sub-Saharan Africa. The quality of anti-malarials in most low-and-middle income countries (LMICs) is affected by several factors including inadequate regulation and limited resources. In this study, the pharmacopeial quality of artemether–lumefantrine (AL) in low and high malaria transmission settings in Uganda was assessed. Methods This was a cross-sectional study conducted among randomly selected private drug outlets. The AL anti-malarials available in drug outlets were purchased using overt method. The samples were screened for quality using visual inspection, weight uniformity, content assay and dissolution tests. The assay test was done using liquid chromatography–mass spectrometry (LC–MS). The samples were considered substandard if the active pharmaceutical ingredient (API) content was outside 90–110% range of the label claim. Dissolution test was conducted following United States Pharmacopoeia (USP) method. Data was analysed using descriptive statistics and presented as means with standard deviations, frequencies, and proportions. Correlation between medicine quality and independent variables was determined using Fisher’s exact test of independence at 95% level of significance. Results A total of 74 AL anti-malarial samples were purchased from high (49/74; 66.2%) and low (25/74; 33.8%) malaria transmission settings. The most common batch of AL was LONART, 32.4% (24/74), with 33.8% (25/74) being ‘Green leaf’. Overall prevalence of substandard quality artemether–lumefantrine was 18.9% (14/74; 95% CI: 11.4–29.7). Substandard quality AL was significantly associated with setting (p = 0.002). A total of 10 samples (13.5%) failed artemether content assay test while, 4 samples (5.4%, 4/74) failed the lumefantrine assay test. One sample from a high malaria transmission setting failed both artemether and lumefantrine assay content test. Of the samples that failed artemether assay test, 90% had low (< 90%) artemether content. All the samples passed visual inspection and dissolution tests. Conclusion Artemether–lumefantrine agents, the recommended first-line treatment for uncomplicated malaria with APIs outside the recommended pharmacopeial content assay limit is common especially in high malaria transmission settings. There is need for continuous surveillance and monitoring of the quality of artemisinin-based anti-malarials across the country by the drug regulatory agency.

Background: Substandard antimalarial agents are a key challenge to effective malaria control and elimination efforts especially in sub-Saharan Africa. The quality of antimalarial agents in most low-and-middle income countries (LMICs) is affected by several factors including inadequate regulation and limited resources. In this study, we assessed the pharmacopeial quality of Artemether-Lumefantrine (AL) in low and high malaria transmission settings in Uganda. Methods: This was a cross-sectional study conducted among randomly selected drug outlets (pharmacies/drug shops). The AL antimalarial agents available in drug outlets were purchased using overt method. The samples were screened for quality using visual inspection, weight uniformity and content assay tests. The assay test was done using Liquid chromatography-mass spectrometry (LC-MS) following International and Unites States Pharmacopoeia (USP) method. The samples were considered substandard if the Active Pharmaceutical Ingredient (API) content was outside 90-110% range of the label claim. Data was analysed using descriptive statistics and presented as means with standard deviations, frequencies, and proportions. Correlation between medicine quality and independent variables was determined using fisher’s exact test of independence at 95% level of significance.Results: A total of 74 AL antimalarial samples were purchased from high (49/74; 66.2%) and low (25/74; 33.8%) malaria transmission settings. The most common batch of AL was LONART, 32.4% (24/74), with 33.8% (25/74) having a ‘Green leaf logo’. Overall, prevalence of substandard quality artemether-lumefantrine was 18.9% (14/74; 95%CI: 11.4-29.7). Substandard quality AL was significantly associated with setting (p=0.002). A total of 10 samples (13.5%) failed artemether content assay while, 4 samples (5.4%, 4/74) had substandard lumefantrine content. One sample from a high malaria transmission setting failed both Artemether and Lumefantrine assay test. Of the samples that failed artemether assay test, majority, 90% had low (<90%) artemether content. Conclusion: Substandard quality AL, the recommended first-line antimalarial agent in treatment of uncomplicated malaria is common especially in high malaria transmission settings. There is need for regular surveillance and monitoring of the quality of artemisinin based antimalarial agents across the country.

Background Malaria treatment is faced with the challenge of access, affordability, availability, and quality of antimalarial medicines. Affordable medicines facility-malaria (AMFm) program and subsequently Co-payment mechanism were developed to help increase access to quality assured Artemisinin Combination Therapies (ACTs) in seven countries in sub-Saharan Africa. We explored through a qualitative study, experience of healthcare personnel on Co-payment mechanism and the implication on access and availability of ACTs in private drug outlets in Uganda. Method All private drug outlets reporting stocking antimalarial agents in low and high malaria transmission settings were purposely selected for the study. In each drug outlet, data was collected from pharmacists/dispensers through key informant interview. The interview covered the following areas, (i) awareness of the co-payment mechanism, (ii) Knowledge of quality assured artemisinin combination therapies (QAACT), (iii) stocking of QAACTs, (iv) dispensing price of QAACTs), and (v) determinants of dispensing price of QAACTs. Data was managed using Atlas.ti and analyzed using framework methodology. Results Data was collected from 25 key informants. Five themes emerged following data analysis, (i) considerations taken while stocking antimalarial agents, (ii) access and purchasing behavior of clients, (iii) antimalarial dispensing, (iv) awareness of QAACT, and (v) awareness of Co-payment mechanism. None of the respondents was aware of Co-payment mechanism and QAACT (green leaf ACT). Duocotecin brand of ACTs (non-QAACT) was the most stocked antimalarial agent. Every seven in ten drug outlet clients request to purchase ACTs without a prescription and preferred buying cheaper brands. Drug outlets stocked and sold both ACT and non-ACT antimalarial agents. Most drug outlet clients cannot afford buying a full dose of an ACT. None of the respondents considered using Co-payment mechanism while stocking ACTs. Conclusion There is lack of awareness of Co-payment mechanism and QAACT among pharmacists/ dispensers close to a decade after its introduction in private sector. There was reportedly no difference in the dispensing price between QAACT and non-QAACT. The dispensing of less than a full dose of ACTs to drug outlet clients is a common practice. The Ministry of Health needs to create awareness through public campaigns on the Co-payment mechanism in the country.