A miniPCR-Duplex Lateral Flow Dipstick Platform for Rapid and Visual Diagnosis of Lymphatic Filariae Infection
Achinya Phuakrod,
Witsaroot Sripumkhai,
Wutthinan Jeamsaksiri,
Pattaraluck Pattamang,
Sumat Loymek,
Paul J. Brindley,
Patsharaporn T. Sarasombath,
Sirichit Wongkamchai
Affiliations
Achinya Phuakrod
Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand
Witsaroot Sripumkhai
Thai Microelectronic Center, National Electronics and Computer Technology Center, Thailand Science Park, Pathum Thani 12110, Thailand
Wutthinan Jeamsaksiri
Thai Microelectronic Center, National Electronics and Computer Technology Center, Thailand Science Park, Pathum Thani 12110, Thailand
Pattaraluck Pattamang
Thai Microelectronic Center, National Electronics and Computer Technology Center, Thailand Science Park, Pathum Thani 12110, Thailand
Sumat Loymek
Office of Disease Prevention and Control, Region 12, Department of Disease Control, The Ministry of Public Health, Songkhla 9000, Thailand
Paul J. Brindley
Immunology & Tropical Medicine & Research Center for Neglected Diseases of Poverty, Department of Microbiology, School of Medicine & Health Sciences, George Washington University, Washington, DC 20037, USA
Patsharaporn T. Sarasombath
Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand
Sirichit Wongkamchai
Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand
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.
