Detection of Wuchereria bancrofti DNA in paired serum and urine samples using polymerase chain reaction-based systems

Abstract

The Global Program for the Elimination of Lymphatic Filariasis (GPELF) aims to eliminate this disease by the year 2020. However, the development of more specific and sensitive tests is important for the success of the GPELF. The present study aimed to standardise polymerase chain reaction (PCR)-based systems for the diagnosis of filariasis in serum and urine. Twenty paired biological urine and serum samples from individuals already known to be positive for Wuchereria bancrofti were collected during the day. Conventional PCR and semi-nested PCR assays were optimised. The detection limit of the technique for purified W. bancrofti DNA extracted from adult worms was 10 fg for the internal systems (WbF/Wb2) and 0.1 fg by using semi-nested PCR. The specificity of the primers was confirmed experimentally by amplification of 1 ng of purified genomic DNA from other species of parasites. Evaluation of the paired urine and serum samples by the semi-nested PCR technique indicated only two of the 20 tested individuals were positive, whereas the simple internal PCR system (WbF/Wb2), which has highly promising performance, revealed that all the patients were positive using both samples. This study successfully demonstrated the possibility of using the PCR technique on urine for the diagnosis of W. bancrofti infection.

Recent advances in nucleic acid-based methods for detection of helminth infections and the perspective of biosensors for future development

“…For example, a two-step semi-nested PCR for amplifying Oesophagostomum bifurcum egg DNA in human faecal samples was shown to achieve a specificity of 100% (Verweij et al ., 2000). In addition, the semi-nested PCR technique showed a 1 000 000-fold increase to the detection limit for W. bancrofti, compared with that of the conventional PCR (Ximenes et al ., 2014).…”

An update on non-invasive urine diagnostics for human-infecting parasitic helminths: what more could be done and how?

“…Unnamed tandem-specific region (Kanjanavas et al, 2005) PCR (Ximenes et al, 2014) 188 bp fragment from SspI repeat sequence PCR (Lucena et al, 1998) Gastrointestinal tract (adult stage) and/ or circulatory system (larval stage)…”

Technetium-99m-Labeled Diethylcarbamazine Citrate (^99mTc-DEC) as a New Diagnostic Agent for Lymphatic Filariasis Detection

“…Therefore, early detection for this case is quite difficult to attain. Laboratory tests for identification of filarial antigens by enzyme-linked immunosorbent assay (ELISA) technique or rapid immunochromatographic card can actually be done, but those techniques are complicated and often give false positive results [5][6][7][8][9][10][11][12]. Diagnosis techniques with higher sensitivity and specificity are still needed .…”