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Background  Lymphatic filariasis caused by  Wuchereria bancrofti  or  Brugia  spp. is a public health problem in developing countries. To monitor bancroftian filariasis infections, Circulating Filarial Antigen (CFA) test is commonly used, but for brugian infections only microfilariae (Mf) microscopy and indirect IgG4 antibody analyses are available. Improved diagnostics for detecting latent infections are required.    Methods  An optimized real-time PCR targeting the brugian  HhaI  repeat was validated with plasma from microfilariae negative Mongolian gerbils (jirds) infected with  B. malayi . Plasma samples from microfilaremic patients infected with  B. malayi  or  W. bancrofti  were used as positive and negative controls, respectively. PCR results of plasma samples from a transmigrant population in a  B. malayi  endemic area were compared to those of life-long residents in the same endemic area; and to IgG4 serology results from the same population. To discriminate between active infections and larval exposure a threshold was determined by correlation and Receiver-Operating Characteristics (ROC) curve analyses.    Results  The PCR detected  HhaI  in pre-patent (56 dpi)  B. malayi  infected jirds and  B. malayi  Mf-positive patients from Central Sulawesi, Indonesia.  HhaI  was also detected in 9/9 elephantiasis patients. In South Sulawesi 87.4% of the transmigrants and life-long residents (94% Mf-negative) were  HhaI  PCR positive. Based on ROC-curve analysis a threshold for active infections was set to >53  HhaI  copies/μl (AUC: 0.854).    Conclusions  The results demonstrate that the  HhaI  PCR detects brugian infections with greater sensitivity than the IgG4 test, most notably in Mf-negative patients ( i.e . pre-patent or latent infections).

Real-time PCR detection of the HhaI tandem DNA repeat in pre- and post-patent Brugia malayi infections: a study in Indonesian transmigrants