Abstract
Rapid diagnosis and parasitemia measurement is crucial for management of malaria. Microscopic examination of peripheral blood (PB) smears is the gold standard for malaria detection. However, this method is labor-intensive. Here, we aimed to develop a completely automated microscopic system for malaria detection and parasitemia measurement. The automated system comprises a microscope, plastic chip, fluorescent dye, and an image analysis program. Analytical performance was evaluated regarding linearity, precision, and limit of detection and was compared with that of conventional microscopic PB smear examination and flow cytometry. The automated microscopic malaria parasite detection system showed a high degree of linearity for Plasmodium falciparum culture (R2 = 0.958, p = 0.005) and Plasmodium vivax infected samples (R2 = 0.931, p = 0.008). Precision was defined as the %CV of the assay results at each level of parasitemia and the %CV value for our system was lower than that for microscopic examination for all densities of parasitemia. The limit of detection analysis showed 95% probability for parasite detection was 0.00066112%, and a high correlation was observed among all three methods. The sensitivity and specificity of the system was both 100% (n = 21/21) and 100% (n = 50/50), respectively, and the system correctly identified all P. vivax and P. falciparum samples. The automated microscopic malaria parasite detection system offers several advantages over conventional microscopy for rapid diagnosis and parasite density monitoring of malaria.
Highlights
Malaria is one of the most dangerous infectious diseases and causes 229 million infections and 409,000 deaths worldwide, according to the World Health Organization (WHO) estimates [1]
Parasitemia of P. falciparum positive patient samples ranged from 0.05% to 4.75% and the parasitemia of P. vivax positive patient samples ranged from 0.03% to 0.22%
Thick and thin peripheral blood (PB) smears were prepared on glass microscopic slides using venous blood collected into K2EDTA tubes (Becton Dickinson, Sunnyvale, CA, USA)
Summary
Malaria is one of the most dangerous infectious diseases and causes 229 million infections and 409,000 deaths worldwide, according to the World Health Organization (WHO) estimates [1]. The WHO recommends malaria diagnostic tests for all patients suspected of malaria infection before treatment administration, thereby prompting an increase in the demand of tests to 1 billion by 2020 [2]. Microscopic examination of Giemsa-stained peripheral blood (PB) smears is considered the gold standard for malaria diagnosis. Parasite density serves as one of the diagnostic criteria for severe malaria infection and its monitoring is important for the diagnosis and treatment of malaria. Parasitemia is one of the essential indicators of the therapeutic effects of antimalarial drugs, and daily blood smear analysis is recommended to document the decrease in parasite density until the absence of parasite on treatment day 7 [7]. Microscopic examination of thick and thin blood smears is the standard method for parasitemia measurement. In thick-smear analysis, parasites and 200 or 500 white blood cells (WBCs) are counted according to the number of parasites found, whereas thin blood smears are evaluated for the percentage of infected red blood cells (RBCs) after counting a minimum of 500 to 2000 RBCs [12]
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