Operational Performance of a Plasmodium falciparum Ultrasensitive Rapid Diagnostic Test for Detection of Asymptomatic Infections in Eastern Myanmar.

In the Greater Mekong Subregion in Southeast Asia, malaria elimination strategies need to target all Plasmodium falciparum parasites, including those carried asymptomatically. More than 70% of asymptomatic carriers are not detected by current rapid diagnostic tests (RDTs) or microscopy. An HRP2-based ultrasensitive RDT (uRDT) developed to improve the detection of low-density infections was evaluated during prevalence surveys within a malaria elimination program in a low-transmission area of eastern Myanmar. Surveys were conducted to identify high-prevalence villages. Two-milliliter venous blood samples were collected from asymptomatic adult volunteers and transported to the laboratory. Plasmodium parasites were detected by RDT, uRDT, microscopy, ultrasensitive qPCR (uPCR), and multiplex enzyme-linked immunosorbent assay (ELISA). The sensitivity, specificity, and predictive positive and negative values of RDT and uRDT were calculated compared to uPCR and ELISA. Parasite and antigen concentrations detected by each test were defined using uPCR and ELISA, respectively. A total of 1,509 samples, including 208 P. falciparum-positive samples were analyzed with all tests. The sensitivity of the uRDT was twofold higher than that of RDT, 51.4% versus 25.2%, with minor specificity loss, 99.5% versus 99.9%, against the combined reference (uPCR plus ELISA). The geometric mean parasitemia detected by uRDT in P. falciparum monospecific infections was 3,019 parasites per ml (95% confidence interval [95% CI], 1,790 to 5,094; n = 79) compared to 11,352 parasites per ml (95% CI, 5,643 to 22,837; n = 38) by RDT. The sensitivities of uRDT and RDT dropped to 34.6% and 15.1%, respectively, for the matched tests performed in the field. The uRDT performed consistently better than RDT and microscopy at low parasitemias. It shows promising characteristics for the identification of high-prevalence communities and warrants further evaluation in mass screening and treatment interventions.

Detection of foci of residual malaria transmission through reactive case detection in Ethiopia.

BACKGROUND: Sub-microscopic and asymptomatic infections could be bottlenecks to malaria elimination efforts in Ethiopia. This study determined the prevalence of malaria, and individual and household-level factors associated with Plasmodium infections obtained following detection of index cases in health facilities in Jimma Zone. METHODS: Index malaria cases were passively detected and tracked in health facilities from June to November 2016. Moreover, family members of the index houses and neighbours located within approximately 200 m from the index houses were also screened for malaria. RESULTS: A total of 39 index cases initiated the reactive case detection of 726 individuals in 116 households. Overall, the prevalence of malaria using microscopy and PCR was 4.0% and 8.96%, respectively. Seventeen (43.6%) of the index cases were from Doyo Yaya kebele, where parasite prevalence was higher. The majority of the malaria cases (90.74%) were asymptomatic. Fever (AOR = 12.68, 95% CI 3.34-48.18) and history of malaria in the preceding 1 year (AOR = 3.62, 95% CI 1.77-7.38) were significant individual-level factors associated with detection of Plasmodium infection. Moreover, living in index house (AOR = 2.22, 95% CI 1.16-4.27), house with eave (AOR = 2.28, 95% CI 1.14-4.55), area of residence (AOR = 6.81, 95% CI 2.49-18.63) and family size (AOR = 3.35, 95% CI 1.53-7.33) were main household-level predictors for residual malaria transmission. CONCLUSION: The number of index cases per kebele may enhance RACD efforts to detect additional malaria cases in low transmission settings. Asymptomatic and sub-microscopic infections were high in the study area, which need new or improved surveillance tools for malaria elimination efforts.

Effective High-Throughput Blood Pooling Strategy before DNA Extraction for Detection of Malaria in Low-Transmission Settings.

In the era of (pre) elimination setting, the prevalence of malaria has been decreasing in most of the previously endemic areas. Therefore, effective cost- and time-saving validated pooling strategy is needed for detection of malaria in low transmission settings. In this study, optimal pooling numbers and lowest detection limit were assessed using known density samples prepared systematically, followed by genomic DNA extraction and nested PCR. Pooling strategy that composed of 10 samples in 1 pool, 20 µl in 1 sample, was optimal, and the parasite density as low as 2 p/µl for both falciparum and vivax infection was enough for detection of malaria. This pooling method showed effectiveness for handling of a huge number of samples in low transmission settings (<9% positive rate). The results indicated that pooling of the blood samples before DNA extraction followed by usual nested PCR is useful and effective for detection of malaria in screening of hidden cases in low-transmission settings.