REAL-TIME PCR WITH HIGH-RESOLUTION MELTING ANALYSIS FOR DIAGNOSIS OF LYMPHATIC FILARIASIS

Lymphatic filariasis (LF), or elephantiasis, is caused by the filarial nematodes Wuchereria bancrofti, Brugia malayi, and B. timori. Over a billion people are at risk of contracting the disease, and about 40 million people suffer from severe disfigurement and disability as a result. Diagnosis is an essential element in the management of the disease, both at the individual patient care level and at the level of disease control in endemic populations. The success of LF elimination programs has lowered the levels of microfilariae in blood samples, which in turn has reduced the sensitivity of the conventional microfilariae detection method. In this study, we used real-time PCR with highresolution melting analysis (HRM) to detect and identify B. malayi and W. bancrofti in human blood samples. The assay can be performed using a single pair of primers targeting the mitochondrial partial 12S rRNA genes of these filarial parasites. Amplification and detection steps can be conducted in the same reaction, and no post-PCR processing is required. After assessing 30 blood samples, the HRM assay exhibited a specific melting temperature for each species, as well as high sensitivity and specificity compared with the microfilaria-detection method. Thus, the assay could be helpful for the diagnosis of LF.