Molecular detection and epidemiological characterization of dengue and malaria coinfection in the Guiriko region, Burkina Faso.

Challenges and strategies in malaria control: A case series from East Azerbaijan Province, 2023-2024.

The co-occurrence of malaria and HIV represents a significant health crisis in sub-Saharan Africa, particularly in Ethiopia, where both diseases are prevalent. This synergistic burden aggravates health inequalities, strains healthcare infrastructure, and challenges socioeconomic stability. Investigating the relationship between these co-infections is crucial for enhancing treatment, prevention, diagnosis, and control strategies for malaria in individuals living with HIV. Therefore, this systematic review and meta-analysis aimed to determine the overall burden of malaria-HIV co-infection, as well as the rates of severe anemia, hyperparasitemia, and CD4 + T-cell counts. A comprehensive literature search was conducted using several electronic databases, including PubMed, Science Direct, Scopus, and Cochrane, as well as search engines like Google Scholar. A random-effects model was used to calculate pooled burden estimates. Subgroup analysis and funnel plots were utilized to evaluate heterogeneity and publication bias, respectively. Of the 429 studies initially reviewed, ten articles met the criteria for inclusion in the final analysis. The pool prevalence estimate of malaria-HIV co-infection was 9% (95% CI 3-15). When grouped by the year of conduct, studies from 2013 to 2025 exhibited the highest malaria burden at 10% (95% CI -4-24), while those conducted from 2002 to 2012 showed the lowest burden at 9% (95% CI 1-16). The overall combined prevalence rates for specific clinical conditions were 31% (95% CI 12-50) for severe anemia, 41% (95% CI -8-90) for hyperparasitemia, and 35% (95% CI 14-56) for low CD4 counts (≤ 200 cells/µL). This meta-analysis found that the overall burden of malaria-HIV co-infection in Ethiopia is 9%.

The ANOFEL/LABAC recommendations aim to standardize the methods used for the biological diagnosis of malaria in France in accordance with the NF EN ISO 15189 standard. Any fever after returning from a malaria-endemic area should raise suspicion of malaria and prompt urgent testing for Plasmodium. Biological diagnosis is based on detecting parasites or their DNA using a sensitive technique, such as rapid molecular biology or a thick smear. If the result is positive, a thin blood smear is performed to identify the parasite species and stages and estimate parasitemia. A rapid diagnostic test (RDT) for detecting plasmodial proteins is a supplementary diagnostic tool and should not be used alone. The result must confirm or rule out malaria, and if malaria is confirmed, it must specify the species involved and the parasitemia. Biologists play an active role in rapidly communicating results and in the medical management of patients. They collaborate with clinicians, interpret results, identify serious cases, and contextualize findings. In the event of indigenous malaria, they participate in mandatory reporting (MR) to the regional health agency (ARS). CNR- or LBMR-qualified malaria laboratories may be consulted for expertise in severe or complex cases. Operator competence and reagent performance control are essential to ensuring the reliability of the results produced.

Malaria remains a significant global health challenge, requiring diagnostic approaches that are rapid, cost-effective, and accurate. The present study proposes a hybrid deep learning framework for malaria diagnosis using thin blood smear cell images obtained from the National Institute of Malaria Research (NIMR), a leading research institution of the Indian Council of Medical Research (ICMR). A total of 15,938 thin blood smear microscopic images were preprocessed to enhance the visibility of cellular structures and suppress background noise. Segmentation was performed using the Otsu thresholding method, which automatically determines the optimal threshold to maximize cell contrast and improve feature extraction. The processed images were then used to train the proposed Hybrid Inception-v3 convolutional neural network (CNN), specifically designed for automated cell classification. To establish the robustness of the proposed CNN, comparative and statistical analyses were performed against conventional diagnostic techniques and other state-of-the-art machine learning models. The proposed hybrid CNN model achieved a training accuracy of 98.9%, sensitivity of 97.3%, specificity of 99.9%, along with high F1 score and area under the curve (AUC) values. These results were obtained using the ICMR-NIMR microscopic image dataset, demonstrating the robustness and generalizability of our approach. The model consistently outperformed other evaluated methods, including state-of-the-art machine learning classifiers, and demonstrated performance comparable to or better than conventional microscopy-based examinations. The hybrid deep learning framework demonstrated robust performance for malaria diagnosis using microscopic images, suggesting potential to reduce reliance on expert microscopists. These findings highlight its potential to improve diagnostic precision and strengthen malaria control efforts, especially in resource-limited settings.

This study set out to describe imported pediatric malaria in the United States over an 8-year period, including patient demographics, clinical outcomes, and risk factors for severe disease. A retrospective descriptive study of pediatric patients treated for malaria at 9 hospitals in the United States from 2016 to 2023 was conducted to analyze patient demographics, clinical outcomes, and risk factors for severe malaria. A total of 171 children were treated across the 9 US hospitals included in this study from 2016 to 2023. Most patients had traveled to West Africa to visit friends and relatives. Fever was the most common symptom recorded (90%), and most reported at least 1 abdominal symptom (66%). Thirty-two percent of patients were diagnosed with severe malaria. No deaths occurred. Delayed diagnoses were common (26%), occurred at similar rates across all hospitals in the study, and were a risk factor for severe malaria. Delayed diagnoses of travel-acquired malaria were common for pediatric patients presenting to hospitals in the United States and are associated with higher risk for severe malaria, which is associated with longer hospitalizations and increased morbidity. Chemoprophylaxis against travel-acquired malaria and prompt diagnosis of imported cases are crucial to improving patient outcomes.

Fever in the returning international traveler presents a diagnostic challenge in the emergency department due to the wide range of potential infectious diseases that may be encountered abroad, many of which may manifest with nonspecific presentations. Malaria remains the most common and life-threatening tropical cause of fever and must be ruled out in all febrile patients returning from malaria-endemic regions. Other key infections to consider in returning travelers include dengue, enteric fever, and leptospirosis. Early recognition and treatment of these diseases can reduce morbidity and mortality and may help contain disease outbreaks. This review presents a structured, systematic approach to the timely evaluation of fever in returning international travelers, including evidence-based recommendations for the diagnosis and treatment of malaria, dengue, enteric fever, and leptospirosis.

Egypt has recently been certified as malaria-free. To sustain this status, the Egyptian Ministry of Health has implemented a policy of centralised malaria treatment at specialised hospitals, provided free of charge to all populations regardless of migrant status. This study examines migrants' malaria care-seeking experiences, the factors influencing their choices, and strategies to improve access. A qualitative study was conducted in Cairo using semi-structured interviews, inviting adult migrants from malaria-endemic countries and with prior malaria experience. Participants were recruited through purposive and convenience sampling. Data were collected in Arabic, transcribed, translated, and analysed thematically. We recruited 21 individuals, all Sudanese; 90.5% were female. Five participants (24%) developed malaria symptoms immediately upon arrival in Egypt; others developed symptoms within six months. Three themes emerged from the analysis: "Malaria-specific healthcare practices," "Determinants of healthcare-seeking behaviour," and "Recommendations." Although participants recognised the severity of malaria and the need for treatment, 19 (90.5%) initially relied on traditional remedies or self-medication without malaria testing. Factors influencing healthcare-seeking behaviour included, at the patient level, a lack of trust in healthcare professionals, reluctance to disclose a malaria diagnosis due to stigma, fears of quarantine, concerns about adverse drug effects, and exposure to negative rumours and misconceptions about treatment. Factors within the healthcare system encompassed malaria services' affordability, accessibility, and perceived quality. Sudanese migrants' malaria care-seeking behaviours are influenced by multiple personal and systemic factors. Improving access requires targeted health education to address misconceptions, raise awareness, and build trust in the healthcare system.

Accurate malaria diagnosis is crucial for effective case management, strong surveillance, and progress toward elimination. However, in highland regions, diagnostic tools are underutilized or yield suboptimal performance. While hematological alterations are frequently observed in malaria, their role remains largely supportive rather than diagnostic. This study aimed to evaluate diagnostic challenges by comparing the performance of HRP2-based rapid diagnostic tests, microscopy, and PCR at Bichena Primary Hospital, Northwest Ethiopia, with hematological profiles examined as supportive indicators to help contextualize diagnostic performance. A facility-based cross-sectional study was conducted between 31/12/2024 to 28/02/2025, with 274 participants enrolled through consecutive sampling. Socio-demographic data were collected using semi-structured questionnaires. The diagnostic evaluation used nested polymerase chain reaction (PCR) (from dried blood spots), microscopy (capillary and venous blood), histidine rich protein 2 (HRP2)-based rapid diagnostic tests (RDTs), and hematological profiling. Data analysis was carried out with Statistical Package for the Social Sciences (SPSS) version 25.0, assessing diagnostic accuracy through sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV), while inter-test agreement was measured using Cohen's Kappa coefficient. Results were summarized in text, figures, and tables. Higher prevalence of Plasmodium infections was detected in 23.4% of participants by PCR, 20.1% by microscopy and 19% by HRP2-antigen-based RDT. The HRP2-antigen based RDT showed lower sensitivity (79.1%), NPV (94.1%), and test accuracy (94.9%) compared to PCR. Similarly, microscopy exhibited high specificity and PPV (100%); however, the sensitivity was 85.9%, indicating that some true positives are missed compared to PCR. Moderate test agreement was observed between PCR and microscopy (κ=0.904; P=0.00) but weak agreement between PCR and RDTs (κ=0.847). Hematological analysis revealed a significantly lower platelet count among PCR-confirmed malaria cases (P<0.05), suggesting a supportive association rather than diagnostic utility. Both HRP2-antigen based RDTs and microscopy demonstrated lower sensitivity compared to PCR. RDTs showed the lowest diagnostic potential for P. falciparum, mixed and even P. vivax infections, this may be due to low parasitemia and possible pfhrp2 deletions. Hematological parameters, particularly platelet count, may serve as complementary indicators to support clinical suspicion but should not replace parasitological or molecular diagnosis. Further investigation of pfhrp2/pfhrp3 deletions is critical to inform the selection of appropriate diagnostic tools in the area.

Plasmodium vivax malaria is a threat to armed forces operating in the Korean Peninsula. Surges in malaria cases in the Republic of Korea entail increased risk. We report 6 cases from a cluster of P. vivax malaria cases in a non-endemic setting in active duty personnel who had redeployed from the Korean Peninsula. Demographics, disease course, and laboratory data were collected on 6 patients diagnosed with Plasmodium vivax malaria at Fort Bliss in 2022. All patients were males, with a mean age of 23 years (range 20-27 years), exposed in the Dagmar North training area in the Gyeonggi province in 2021, with an average time from symptom onset to diagnosis of 57 days (8-121 days). All had uncomplicated malaria. Laboratory findings included hemoglobin of 12.1g/dL (range 8.1-14.2 g/dL), platelets of 123 × 103/µL (range 40-171 × 103/µL), and parasitemia <1% (0.1%-0.9%) with diagnosis on peripheral smear and/or rapid antigen testing. No cases received chemoprophylaxis. Patients were treated with artemether/lumefantrine and primaquine. Clearance of parasitemia on peripheral smear was seen after an average of 2.8 days (range 2-4 days). Changing vector ecology and increased tempo of training exercises in the Dagmar North Region near the Demilitarized Zone are hypothesized to contribute to this cluster of cases. Malaria diagnosis post-deployment in non-endemic regions is challenging due to the long latency of illness onset and requires clinician vigilance. Preventive measures of pre-deployment should also be emphasized.

Early and accurate detection of malaria is essential for appropriate case management, minimizing unnecessary treatment, and supporting surveillance efforts crucial for disease control and elimination. Microscopy has long been the reference method for malaria diagnosis. Rapid diagnostic tests (RDTs) have revolutionised point-of-care diagnosis of malaria, with billions of units distributed globally over the past decade. However, ensuring their reliability necessitates rigorous quality assurance measures. Inaccurate test performance can lead to misdiagnosis, inappropriate treatment, increased mortality, and the potential emergence of drug-resistant strains that can lead to delay in achieving the target of malaria elimination. Strengthening quality assurance protocols is crucial to maintain the accuracy of malaria diagnostics, enhancing disease surveillance, and bolstering global malaria elimination initiatives. This paper focuses on the role of quality assurance in optimising the performance and reliability of malaria RDTs. By implementing enhanced quality assurance frameworks, diagnostic accuracy can be safeguarded, toward malaria eradication.

The Chikungunya virus (CHIKV) is an arbovirus transmitted by hematophagous arthropods, widely distributed in tropical and subtropical regionsAQ1. In Brazil, its introduction in 2014 marked the beginning of a significant expansion, becoming a major public health problem. This work consists of a cross-sectional study conducted from October 2024 to July 2025, in which 830 samples from patients with acute febrile illness and negative diagnosis for malaria were evaluatedAQ2. The samples were tested for dengue, Zika, and chikungunya and subjected to duplex RT-qPCR to detect Mayaro and Oropouche. Of the samples analyzed, 5,18% (43/830) tested positive for some arbovirus investigated in this cohort, of which 53,49% (23/43) were detectable for CHIKV. The phylogenetic analysis showed that all samples belonged to the East/Central/South African (ECSA) genotype, demonstrating its circulation in the region. The findings reinforce the need for continuous molecular surveillance to monitor the circulation of medically important arboviruses in the Amazon region.

Although clinical malaria occurs far less frequently in newborns and young infants aged <6months than in older children, its importance and public health relevance should not be ignored. This population is often perceived to have protection against malaria, and making a diagnosis of malaria in this age group can be challenging, as the symptoms generally overlap with those of many other common conditions in infancy, possibly leading to misdiagnosis or treatment delay. This narrative review synthesises current evidence, explores existing treatment recommendations, and highlights practice gaps in malaria management in children aged <6months (or weighing <5kg). A literature search was conducted using PubMed, EMBASE, and Google Scholar and supplemented with World Health Organization and region-specific malaria treatment guidelines. Data on malaria prevalence in young infants are highly variable and heterogeneous, reflecting differences in the methodology used, diagnostic criteria and treatment approaches. Evidence regarding the safety and efficacy of available antimalarial therapies in this population is limited, and until recently, no antimalarial agents were licensed for neonates or infants weighing <4.5kg. Consequently, current recommendations may not adequately address the needs of this vulnerable group. Given the variability in the guidelines across different countries, there is an urgent need to make specific policy and practice amendments that can enhance understanding, bridge existing knowledge gaps and harmonise treatment approaches for at-risk populations worldwide.

This study aimed to describe the cross-reactivity of the Abbott Bioline Malaria Ag P.f/P.f/P.v rapid diagnostic test, designed to detect Plasmodium falciparum and P. vivax, with other Plasmodium species, based on the detection of parasite lactate dehydrogenase enzymes. Blood samples from 40 patients with positive thick and thin blood smear results were analysed: 10 infected with P. falciparum, 10 with P. vivax, 15 with P. malariae and five with P. ovale. Parasitemia was quantified as parasites/μL of blood. All samples were tested using the Abbott Bioline Malaria Ag P.f/P.f/P.v RDT, and species identification was confirmed by nested PCR. To compare test line intensities and assess potential nonspecific reactions, images of the RDT lines were analysed using ImageJ software. For statistical analyses, PCR was considered the gold standard. Sensitivity, specificity and accuracy were calculated using MedCalc 2026. Fisher's exact test was applied using GraphPad/Quick Calcs to compare the specificity of the anti-Pf-pLDH antibodies, with a significance level of 0.05 (p < 0.05). The RDT accurately detected all P. falciparum and P. vivax samples. However, 12 of 14 P. malariae samples yielded positive results on the T2 line, intended to be specific for Pf-pLDH, as did four P. ovale isolates. The specificity of the anti-Pf-pLDH monoclonal antibodies against P. malariae was only 14.29% (p < 0.05). ImageJ analysis showed that the T2 line signal intensity in P. malariae-infected samples exceeded that observed in P. falciparum-infected samples. This study provides the first evidence of cross-reactivity of anti-Pf-pLDH monoclonal antibodies in the Abbott Bioline Malaria Ag P.f/P.f/P.v RDT with P. malariae and P. ovale isolates. Moreover, highlights the imperative need to validate alternative biomarkers and novel point-of-care platforms, to enhance the rapid and accurate diagnosis of malaria, ensure species-specific treatment and ultimately support effective malaria control and elimination strategies.

Malaria remains a major global health challenge, particularly in low-resource settings where diagnostic infrastructure is limited. The gold standard diagnostic is manual light microscopy which is slow, labor-intensive, and dependent on expert interpretation limiting the management of malaria. Here, we present a magneto-optical (MO) imaging approach that exploits the magnetic anisotropy and optical dichroism of hemozoin crystals to achieve label-free, quantitative detection of malaria infection within a conventional polarizing microscope. In our implementation, an external magnetic field induces torque on the paramagnetic hemozoin crystals, aligning their long axes along the field vector. This alignment modulates the optical anisotropy of the specimen, producing a measurable field-dependent change in transmitted polarization contrast. Unlike previously reported standalone magneto-optical devices, which rely on bulk optical scattering or transmission modulation, our approach integrates MO directly into a conventional polarizing microscope setup, allowing simultaneous imaging and quantitative MO signal analysis at the microscopic scale. By integrating ratiometric intensity analysis and threshold-based segmentation metrics, we advance malaria imaging beyond purely observational assessment, enabling a quantitative correlation between MO signal strength and hemozoin concentration, and thus, with parasite developmental stage. These metrics are also well-suited for automated malaria diagnostics, as they provide stable, objective parameters that can be reliably computed without expert intervention. Moreover, because microscopy expertise is already well established in many malaria-endemic regions, embedding this approach within the familiar framework of microscopy offers a practical and rapid pathway for integrating the technique into routine malaria diagnosis.

Effective malaria case management relies on accurate diagnosis and appropriate treatment. However, rapid diagnostic test (RDT) performance varies, and presumptive treatment remains common. We quantified malaria misdiagnosis and inappropriate treatment and assessed adherence to treatment guidelines in a high-burden setting. We leveraged samples collected during a cohort study of children and adults from 2015 to 2022 across seven sites in Kinshasa Province, Democratic Republic of the Congo (DRC). We estimated the cumulative incidence and prevalence of false-positive and false-negative RDT results using PCR as the gold standard and identified correlates of misdiagnosis and inappropriate treatment using Kaplan Meier and generalized estimating equations. Among 2,269 participants, the one-year cumulative incidences of false-positive and false-negative RDTs were 23.1% and 17.8%, respectively; by seven years, these rose to 77.7% and 83.4%. Misdiagnosis (false-positive or -negative RDT) was associated with age, recent treatment, transmission intensity, and parasite density -factors that could inform refinements to treatment guidelines. About 58% of RDT-negative participants received antimalarial treatment at a clinic visit, while 4% of RDT-positive participants did not. Fever significantly reduced adherence to test-and-treat guidelines. Most participants experienced malaria misdiagnosis by RDT over the course of the study, underscoring the limitations of currently used RDTs and opportunities for improved clinical decision-making. While RDTs remain essential for malaria case management, clinicians should consider contextual factors when interpreting results. Efforts to improve adherence to guidelines, which require a positive diagnostic test result before treatment, and management of non-malarial febrile illness are needed in the DRC and similar high-burden settings.

Plasmodium falciparum (P. falciparum) constitutes the main malaria parasite in Niger. Little is known about other malaria parasite species in the country. This study documented the prevalence and spatial distribution of Plasmodium species in the Niger Republic based on different epidemiological zones of the country. This was a cross-sectional study conducted during the high malaria transmission period (September to November 2022). Blood sample was collected on filter paper, from patients with fever. Plasmodium species were identified by RDTs, thick smear, and polymerase chain reaction (PCR). The study population included a total of 5251 patients from whom 5096 were analyzed. Among the analyzed patients, 1460 were in the hypo-endemic epidemiological area, 878 in the meso-endemic epidemiological area and 2758 in the hyperendemic area. Among the 304 samples collected via PCR, 55% (166/304) belonged to the genus Plasmodium. The species falciparum represented 90% (149/166) compared to 5% (9/166) Ovale; 3% (5/166) malariae and 2% (3/166) vivax. The vivax was identified in a hypo-endemic area. This study reveals a strong predominance of P. falciparum in the three epidemiological area of malaria in Niger. The findings highlight the circulation of P. vivax in hypo-endemic area. The results would help the National Malaria Control Program (NMCP) to better orient and adapt its malaria diagnosis and management strategy in Niger.

Unveiling the overlooked burden of malaria misdiagnosis using lamp-based re-evaluation of routine malaria diagnosis in health centers: implications for public health and clinical practice in Northwest Ethiopia.

In the Greater Mekong Subregion, militaries constitute a critical but often underserved malaria transmission reservoir, given their high mobility, deployment to endemic areas, and frequent exposure to vectors. In Thailand, the Royal Thai Army (RTA) is a key risk population, yet their malaria practices, perceptions, and the scale and scope of coordination with the Ministry of Public Health (MoPH) are not well understood. A joint military-civilian research team conducted a rapid formative assessment in Sisaket Province, a persistent transmission hotspot, to characterize the unique RTA risk profile and identify opportunities to strengthen RTA-MoPH coordination on malaria elimination efforts. Using a mixed-methods design, the research team conducted a five-year (2016-2020) retrospective analysis of Sisaket's malaria case and program response data, alongside 16 focus group discussions (FGDs) and 17 key informant interviews (KIIs) with RTA and MoPH respondents across all military ranks and health system levels in the province's three highest-burden districts (Kantharalak, Khun Han, Phu Sing). Qualitative data were collected between December 2021-January 2022 and thematically analyzed using an inductive content analysis approach. Quantitative data were descriptively analyzed using statistical methods to characterize the RTA malaria risk profile and identify and compare recent trends in program response between military and non-military populations in Sisaket Province. 432 military malaria cases were reported in Sisaket between 2016 and 2020, accounting for 18% of the province's total 2425 cases (with the proportion ranging from 14 to 30% annually). All military malaria cases were male, with a median age of 28years. 96% of military cases were diagnosed and treated at MoPH facilities, with 41% of cases classified as indigenous and 40% as imported from abroad. Qualitative data were collected from a total of 116 respondents (72 RTA, 44 MoPH) through 16 FGDs and 17 KIIs. Malaria prevention and treatment practices and perceptions among soldiers were largely consistent across the three study districts. RTA-MoPH coordination occurred at all levels through both formal and informal channels and was reported to be improving and expanding by some respondents. Though several areas could benefit from increased collaboration: patient follow-up, border control efforts, vector control, and further capacitation of military medics. The RTA and MoPH both expressed strong interest in strengthening military-civilian coordination; though to better enable this, several identified challenges may need to be addressed, including restricted MoPH access to military sites, lack of continuity in relationships due to frequent RTA rotations, communication constraints, and broader differences in military versus civilian operating procedures. Strengthening RTA-MoPH coordination towards successfully interrupting malaria transmission in Thailand may require formalizing and standardizing some joint operating procedures, increasing communication and military-civilian touchpoints, and further capacitating the RTA to carry out malaria prevention, diagnosis and treatment, and patient follow-up activities. For Thailand to achieve its goal of nationwide malaria elimination, it is crucial that the RTA as a high-risk population be fully engaged in the country's malaria control and response efforts.

BackgroundAlthough rapid diagnostic tests (RDTs) are widely used for malaria diagnosis, they have notable limitations. Blood smear (BS) microscopy remains the gold standard, yet its reliability in public health facilities (HFs) across malaria-endemic regions of Kenya can be compromised by limited infrastructure, technical capacity, and quality assurance.MethodsWe assessed the quality of malaria microscopy in 29 HFs in Siaya County, western Kenya, from January–July 2024 by evaluating the concordance of routine HF BS results with expert microscopy. We evaluated the availability and quality of reagents, standard operating procedures, and infrastructure using the National Malaria Control Program (NMCP) technical supervision checklist which follows WHO-certified microscopy standards. Up to 60 participant slides were randomly selected over three visits and two slides were prepared for each participant. Slide 1 was prepared and read on-site as per routine HF practice, then re-examined by an expert microscopist using the HF microscope and again at the Kenya Medical Research Institute (KEMRI). Slide 2 was stained and read at KEMRI, serving as the gold standard. We evaluated factors and characteristics associated with accurate diagnosis using logistic regression.ResultsOf the 1,494 blood smears examined, 501 (34%) were positive. Concordance between routine microscopy and expert re-reading was 91% (1,289/1,414), ranging from 55% (6/11) to 100% (60/60) across health facilities. Percent agreement between HF slide 1 and slide 2 was 86% (1,276/1,485), with a range from 55% (6/11) to 98% (39/40) by HF. Compared to slide 2, sensitivity and specificity of HF results was 76% and 92%, respectively, resulting in undertreatment of 24% and overtreatment of 8% of patients. Those with parasitemia 1–100 p/μL had lower odds of accurate diagnosis (OR = 0.12; 95% CI: 0.05–0.29; p < 0.001) while specimens with parasite densities > 10,000 p/μL had higher odds of accurate diagnosis (OR = 6.08; 95% CI: 2.22–25.1; p = 0.002), indicating a positive association between parasite density and diagnostic accuracy. Six (21%) of HFs had poor quality BS with debris or contamination.ConclusionsWhile overall concordance was high, the variability in results by HF, limited accuracy at low parasite densities, and challenges with required infrastructure highlight the need for ongoing malaria microscopy quality assurance to ensure proper case management.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12936-026-05886-0.