Abstract
Introduction Accurate detection of filarial parasites in humans and vectors is essential for the implementation and evaluation of Global and National Programs to eliminate lymphatic filariasis. Immunological methods to detect infection are available; however, cross-reactivity issues have been reported in most of them. Nucleic acid-based molecular assays offer high levels of specificity and sensitivity and can be used to detect the infections. Methods In this study, we evaluated loop-mediated isothermal amplification (LAMP) tests to amplify Wuchereria bancrofti DNA in patients' blood. The amplicons were tested by both pH-sensitive dyes for enhanced visual detection and agarose gel electrophoresis. A closed-tube LAMP assay was also evaluated. Cohen's Kappa statistics was used for statistical analysis of the assays. 125 patients consented for blood sampling which were used for clinical analysis of LAMP assays with the PCR method used as the “gold standard.” Results The sensitivity of the evaluated Wuchereria bancrofti LAMP was 92.3%, with a specificity of 97.3% and kappa statistics value of 0.84, which is in a strong agreement. Conclusion In this study, LAMP assays coupled with fluorescence dye detection have been found to be suitable for diagnosis and monitoring of Wuchereria bancrofti infections in the Kenyan population.
Highlights
Accurate detection of filarial parasites in humans and vectors is essential for the implementation and evaluation of Global and National Programs to eliminate lymphatic filariasis
In 2000, the Global Program to Eliminate Lymphatic filariasis (LF) (GPELF) was launched [7] and it was estimated that 5–6 rounds of mass drug administration (MDA) with ivermectin or diethylcarbamazine and albendazole were required to eliminate the disease [5]
The results described here show the amplification of W. bancrofti DNA using loop-mediated isothermal amplification (LAMP) primers targeting the Ssp 1 repeat sequence
Summary
Accurate detection of filarial parasites in humans and vectors is essential for the implementation and evaluation of Global and National Programs to eliminate lymphatic filariasis. LAMP assays coupled with fluorescence dye detection have been found to be suitable for diagnosis and monitoring of Wuchereria bancrofti infections in the Kenyan population. Lymphatic filariasis (LF) is a chronic parasitic disease of public health and socioeconomic significance in tropical and subtropical countries. It is caused by the filarial worms of Wuchereria bancrofti, Brugia malayi, or B. timori species. The disease is transmitted by five genera of mosquitoes, Culex, Aedes, Anopheles, Mansonia, and Ochlerotatus [2], making it the most common vector-transmitted parasitic infection after malaria [3, 4] It is the second leading cause of permanent and long-term disability worldwide after eye blindness [5]. In 2018, 893 million people in 49 countries were living in areas that required preventive chemotherapy to stop the spread of the infections [8]
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