A TaqMan-based multiplex real-time PCR assay for the simultaneous detection of Wuchereria bancrofti and Brugia malayi

• Published in: Molecular and biochemical parasitology Vol. 189; no. 1-2; pp. 33 - 37
• Main Authors: Pilotte, N., Torres, M., Tomaino, F.R., Laney, S.J., Williams, S.A.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.05.2013
• Subjects: Animals; Blood - parasitology; Brugia; Brugia malayi; Brugia malayi - genetics; Brugia malayi - isolation & purification; Culicidae; Culicidae - parasitology; Diagnosis; disease surveillance; disease transmission; DNA; DNA, Helminth - genetics; DNA, Helminth - isolation & purification; Elephantiasis, Filarial - diagnosis; Elephantiasis, Filarial - parasitology; Filariasis; Humans; labor; Molecular Diagnostic Techniques - economics; Molecular Diagnostic Techniques - methods; Multiplex PCR; Multiplex Polymerase Chain Reaction - economics; Multiplex Polymerase Chain Reaction - methods; parasites; parasitology; Parasitology - economics; Parasitology - methods; quantitative polymerase chain reaction; Real-time PCR; Real-Time Polymerase Chain Reaction - economics; Real-Time Polymerase Chain Reaction - methods; Sensitivity and Specificity; Wuchereria bancrofti; Wuchereria bancrofti - genetics; Wuchereria bancrofti - isolation & purification; Multiplex PCR; Real-time PCR; Filariasis; Brugia malayi; mf; Wuchereria bancrofti; MDA; GPELF; LDR; Diagnosis
• ISSN: 0166-6851; 1872-9428
• DOI: 10.1016/j.molbiopara.2013.05.001

•We describe a multiplex real-time PCR assay to detect Wuchereria bancrofti and Brugia malayi.•The assay is comparably sensitive to the established singleplex detection assays.•Detection of DNA extracted from mosquito pools or human bloodspots is possible.•The assay provides significant reagent and labor cost savings. With the Global Program for the Elimination of Lymphatic Filariasis continuing to make strides towards disease eradication, many locations endemic for the causative parasites of lymphatic filariasis are realizing a substantial decrease in levels of infection and rates of disease transmission. However, with measures of disease continuing to decline, the need for time-saving and economical molecular diagnostic assays capable of detecting low levels of parasite presence is increasing. This need is greatest in locations co-endemic for both Wuchereria bancrofti and Brugia parasites because testing for both causative agents individually results in significant increases in labor and reagent costs. Here we describe a multiplex, TaqMan-based, real-time PCR assay capable of simultaneously detecting W. bancrofti and Brugia malayi DNA extracted from human bloodspots or vector mosquito pools. With comparable sensitivity to established singleplex assays, this assay provides significant cost and labor savings for disease monitoring efforts in co-endemic locations.