Multiplex Polymerase Chain Reaction for Detection of Dirofilaria immitis (Filariidea: Onchocercidae) and Wuchereria bancrofti (Filarioidea: Dipetalonematidae) in Their Common Vector Aedes polynesiensis (Diptera: Culicidae)

Abstract

In French Polynesia, Aedes polynesiensis (Marks) is the vector of the human filarial parasite Wuchereria bancrofti (Cobbold) and dog heartworm, Dirofilaria immitis (Leidy). A multiplex polymerase chain reaction (PCR) assay was designed to screen pools of field-collected Ae. polynesiensis for the presence of both parasites simultaneously using primers specific for each parasite. The sensitivity of detection on purified DNA was 1 and 10 pg, equivalent to 0.1 and 1 L3 larva per pool for W. bancrofti and D. immitis, respectively. Codetection was performed at an hybridization temperature of 58°C to avoid competition between heterologous DNA and primers that was observed at 55°C. In addition, D. immitis was detected by PCR in the blood of infected dogs.