Multiplex polymerase chain reaction for detection of Dirofilaria immitis (Filariidea: Onchocercidae) and Wuchereria bancrofti (Filarioidea: Dipetalonematidae) in their common vector Aedes polynesiensis (Diptera: Culicidae).

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 degrees C to avoid competition between heterologous DNA and primers that was observed at 55 degrees C. In addition, D. immitis was detected by PCR in the blood of infected dogs.