A miniPCR-Duplex Lateral Flow Dipstick Platform for Rapid and Visual Diagnosis of Lymphatic Filariae Infection.

Achinya Phuakrod,Wutthinan Jeamsaksiri,Sumat Loymek,Sirichit Wongkamchai,Witsaroot Sripumkhai,Patsharaporn T Sarasombath,Paul J Brindley,Pattaraluck Pattamang

doi:10.3390/diagnostics11101855

Abstract

Lymphatic filariasis (LF) is a neglected major tropical disease that is a leading cause of permanent and long-term disability worldwide. Significant progress made by the Global Programme to Eliminate Lymphatic Filariasis (GPELF) has led to a substantial decrease in the levels of infection. In this limitation, DNA detection of lymphatic filariae could be useful due to it capable of detecting low level of the parasites. In the present study, we developed a diagnostic assay that combines a miniPCR with a duplex lateral flow dipstick (DLFD). The PCR primers were designed based on the HhaI and SspI repetitive noncoding DNA sequences of Brugia malayi and Wuchereria bancrofti, respectively. The limits of detection and crossreactivity of the assay were evaluated. In addition, blood samples were provided by Thais living in a brugian filariasis endemic area. The miniPCR-DLFD assay exhibited a detection limit of 2 and 4 mf per milliliter (mL) of blood for B. malayi as well as W. bancrofti, respectively, and crossamplification was not observed with 11 other parasites. The result obtained from the present study was in accordance with the thick blood smear staining for the known cases. Thus, a miniPCR-DLFD is an alternative tool for the diagnosis of LF in point-of-collection settings with a modest cost (~USD 5) per sample.

Highlights

Lymphatic filariasis (LF) caused by W. bancrofti, B. malayi and B. timori, is one of the most neglected tropical diseases
According to the Global Programme to Eliminate Lymphatic Filariasis (GPELF) established by the World Health Organization (WHO), a mass drug administration (MDA) was given to residents of LF endemic areas [4]
The present study proposed for the first time to develop a miniPCR assay coupled with a duplex lateral flow dipstick (DLFD) for rapid and visual detection of both B. malayi and W. bancrofti DNA in human blood samples

Summary

Introduction

Lymphatic filariasis (LF) caused by W. bancrofti, B. malayi and B. timori, is one of the most neglected tropical diseases. An immunochromatographic test (ICT) for the detection of W. bancrofti-circulating antigen is available commercially [8]. Several articles have reported tests that can detect circulating antigens from B. malayi in human blood. Those tests have not yet been independently verified, and none are commercially available [3,12,13,14,15,16]. Antifilarial IgG4 antibody detection has been developed [6,7,17], and it is a useful addition to the limited array of brugian filariasis diagnostic tools available. Antibody tests cannot distinguish between bancroftian and brugian filariasis, limiting their usefulness in areas where these infections are coendemic. In several countries that have completed multiple rounds of mass drug administration, dramatic reductions in both microfilaremia and antigenemia levels have been observed [18]

Methods

Results

Discussion

Conclusion