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

Albers, A., Sartono, E., Wahyuni, S., Yazdanbakhsh, M., Maizels, R. M. , Klarmann-Schulz, U., Pfarr, K. and Hoerauf, A. (2014) Real-time PCR detection of the HhaI tandem DNA repeat in pre- and post-patent Brugia malayi infections: a study in Indonesian transmigrants. Parasites and Vectors, 7, 146. (doi: 10.1186/1756-3305-7-146) (PMID:24685183) (PMCID:PMC4021971)

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Abstract

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).

Item Type:Articles
Additional Information:This work was supported by the European Union, European Commission, INCO programme [contract No. 32321, FP6-INCO]; the Netherlands Foundation for the Advancement of Tropical Research [WOTRO, W93-266]; and intramural funding from University Hospital of Bonn [BONFOR to AA].
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Maizels, Professor Rick
Authors: Albers, A., Sartono, E., Wahyuni, S., Yazdanbakhsh, M., Maizels, R. M., Klarmann-Schulz, U., Pfarr, K., and Hoerauf, A.
College/School:College of Medical Veterinary and Life Sciences > School of Infection & Immunity
Journal Name:Parasites and Vectors
Publisher:BMC
ISSN:1756-3305
ISSN (Online):1756-3305
Copyright Holders:Copyright © 2014 Albers et al.
First Published:First published in Parasites and Vectors 7:146
Publisher Policy:Reproduced in accordance with the publisher copyright policy

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