title: Detection and quantification of Wuchereria bancrofti and Brugia malayi DNA in blood samples and mosquitoes using duplex droplet digital polymerase chain reaction.
abstract: Lymphatic filariasis, a mosquito-borne disease, is still a major public health problem in tropical and sub-tropical countries. Effective diagnostic tools are required for identification of infected individuals, for epidemiological assessment, and for monitoring of control programs. A duplex droplet digital polymerase chain reaction (ddPCR) was conducted to differentiate and quantify Wuchereria bancrofti DNA by targeting the long DNA repeat (LDR) element and Brugia malayi DNA by targeting the HhaI element in blood samples and mosquito vectors. The analytical sensitivity and specificity were evaluated. Our results indicated that the duplex ddPCR assay could differentiate and quantify W. bancrofti and B. malayi DNA from blood samples and mosquitoes. DNA from a single larva in 50 μl of a blood sample, or in one mosquito vector, could be detected. The analytical sensitivity and specificity for W. bancrofti are both 100 %. Corresponding values for B. malayi are 100 and 98.3 %, respectively. Therefore, duplex ddPCR is a potential tool for simultaneous diagnosis and monitoring of bancroftian and brugian filariasis in endemic areas.
authors.affiliation: Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, Thailand.
authors.forename: Jurairat
authors.initials: J
authors.lastname: Jongthawin
authors.affiliation: Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, Thailand.|Department of Parasitology, Khon Kaen University, Khon Kaen, Thailand.
authors.forename: Pewpan M
authors.initials: PM
authors.lastname: Intapan
authors.affiliation: Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, Thailand.|Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
authors.forename: Viraphong
authors.initials: V
authors.lastname: Lulitanond
authors.affiliation: Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, Thailand.|Department of Parasitology, Khon Kaen University, Khon Kaen, Thailand.|Faculty of Medicine, Mahasarakham University, Mahasarakham, Thailand.
authors.forename: Oranuch
authors.initials: O
authors.lastname: Sanpool
authors.affiliation: Faculty of Medicine, Mahasarakham University, Mahasarakham, Thailand.
authors.forename: Tongjit
authors.initials: T
authors.lastname: Thanchomnang
authors.affiliation: Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, Thailand.|Department of Parasitology, Khon Kaen University, Khon Kaen, Thailand.
authors.forename: Lakkhana
authors.initials: L
authors.lastname: Sadaow
authors.affiliation: Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, Thailand. wanch_ma@kku.ac.th.|Department of Parasitology, Khon Kaen University, Khon Kaen, Thailand. wanch_ma@kku.ac.th.
authors.forename: Wanchai
authors.initials: W
authors.lastname: Maleewong
mesh_terms: D000818:Animals; D017178:Brugia malayi / Q000235:genetics*; D002415:Cats; D009033:Culicidae / Q000469:parasitology*; D018509:DNA, Helminth / Q000032:analysis*; D004285:Dogs; D004605:Elephantiasis, Filarial; D006801:Humans; D007814:Larva; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014958:Wuchereria bancrofti / Q000235:genetics*
chemical_list: D018509:DNA, Helminth
keywords: Brugia malayi; Detection; Duplex droplet digital polymerase chain reaction; Lymphatic filariasis; Mosquito vectors; Wuchereria bancrofti