A single fluorescence-based LAMP reaction for identifying multiple parasites in mosquitoes

Area of Science:

• Molecular Biology
• Parasitology
• Vector-borne Diseases

Background:

• Co-endemic distribution of vector-borne diseases like malaria and lymphatic filariasis poses significant public health challenges.
• Simultaneous detection of multiple pathogens in vectors is crucial for effective disease control programs.

Purpose of the Study:

• To develop a multiplex amplification method for simultaneous detection of Plasmodium berghei and Dirofilaria immitis in mosquitoes.
• To evaluate the sensitivity and specificity of the developed LAMP assay for identifying parasite-bearing mosquitoes.

Main Methods:

• Utilized Loop-mediated Isothermal Amplification (LAMP) with fluorescently labeled primers.
• Targeted the SPECT2 gene of P. berghei and the cytochrome oxidase subunit I gene of D. immitis.
• Developed a multiplex assay capable of distinguishing individual parasite species within mosquito samples.

Main Results:

• The multiplex LAMP assay demonstrated high sensitivity, detecting as few as 100 P. berghei-infected red blood cell-equivalents or one D. immitis microfilaria.
• The assay successfully identified individual parasite species when a mixture of primer sets was used.
• The method proved specific for the targeted parasite genes.

Conclusions:

• The multiplex LAMP assay is a sensitive and specific tool for identifying parasite-carrying mosquitoes in co-endemic regions.
• This technique can enhance the efficiency and effectiveness of arthropod-borne disease elimination programs.
• The assay facilitates simultaneous detection of multiple vector-borne pathogens.