Cargando…

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

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Albers, Anna, Sartono, Erliyani, Wahyuni, Sitti, Yazdanbakhsh, Maria, Maizels, Rick M, Klarmann-Schulz, Ute, Pfarr, Kenneth, Hoerauf, Achim
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4021971/
https://www.ncbi.nlm.nih.gov/pubmed/24685183
http://dx.doi.org/10.1186/1756-3305-7-146
_version_ 1782316326847512576
author Albers, Anna
Sartono, Erliyani
Wahyuni, Sitti
Yazdanbakhsh, Maria
Maizels, Rick M
Klarmann-Schulz, Ute
Pfarr, Kenneth
Hoerauf, Achim
author_facet Albers, Anna
Sartono, Erliyani
Wahyuni, Sitti
Yazdanbakhsh, Maria
Maizels, Rick M
Klarmann-Schulz, Ute
Pfarr, Kenneth
Hoerauf, Achim
author_sort Albers, Anna
collection PubMed
description 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).
format Online
Article
Text
id pubmed-4021971
institution National Center for Biotechnology Information
language English
publishDate 2014
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-40219712014-05-16 Real-time PCR detection of the HhaI tandem DNA repeat in pre- and post-patent Brugia malayi infections: a study in Indonesian transmigrants Albers, Anna Sartono, Erliyani Wahyuni, Sitti Yazdanbakhsh, Maria Maizels, Rick M Klarmann-Schulz, Ute Pfarr, Kenneth Hoerauf, Achim Parasit Vectors Research 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). BioMed Central 2014-03-31 /pmc/articles/PMC4021971/ /pubmed/24685183 http://dx.doi.org/10.1186/1756-3305-7-146 Text en Copyright © 2014 Albers et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Albers, Anna
Sartono, Erliyani
Wahyuni, Sitti
Yazdanbakhsh, Maria
Maizels, Rick M
Klarmann-Schulz, Ute
Pfarr, Kenneth
Hoerauf, Achim
Real-time PCR detection of the HhaI tandem DNA repeat in pre- and post-patent Brugia malayi infections: a study in Indonesian transmigrants
title Real-time PCR detection of the HhaI tandem DNA repeat in pre- and post-patent Brugia malayi infections: a study in Indonesian transmigrants
title_full Real-time PCR detection of the HhaI tandem DNA repeat in pre- and post-patent Brugia malayi infections: a study in Indonesian transmigrants
title_fullStr Real-time PCR detection of the HhaI tandem DNA repeat in pre- and post-patent Brugia malayi infections: a study in Indonesian transmigrants
title_full_unstemmed Real-time PCR detection of the HhaI tandem DNA repeat in pre- and post-patent Brugia malayi infections: a study in Indonesian transmigrants
title_short Real-time PCR detection of the HhaI tandem DNA repeat in pre- and post-patent Brugia malayi infections: a study in Indonesian transmigrants
title_sort real-time pcr detection of the hhai tandem dna repeat in pre- and post-patent brugia malayi infections: a study in indonesian transmigrants
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4021971/
https://www.ncbi.nlm.nih.gov/pubmed/24685183
http://dx.doi.org/10.1186/1756-3305-7-146
work_keys_str_mv AT albersanna realtimepcrdetectionofthehhaitandemdnarepeatinpreandpostpatentbrugiamalayiinfectionsastudyinindonesiantransmigrants
AT sartonoerliyani realtimepcrdetectionofthehhaitandemdnarepeatinpreandpostpatentbrugiamalayiinfectionsastudyinindonesiantransmigrants
AT wahyunisitti realtimepcrdetectionofthehhaitandemdnarepeatinpreandpostpatentbrugiamalayiinfectionsastudyinindonesiantransmigrants
AT yazdanbakhshmaria realtimepcrdetectionofthehhaitandemdnarepeatinpreandpostpatentbrugiamalayiinfectionsastudyinindonesiantransmigrants
AT maizelsrickm realtimepcrdetectionofthehhaitandemdnarepeatinpreandpostpatentbrugiamalayiinfectionsastudyinindonesiantransmigrants
AT klarmannschulzute realtimepcrdetectionofthehhaitandemdnarepeatinpreandpostpatentbrugiamalayiinfectionsastudyinindonesiantransmigrants
AT pfarrkenneth realtimepcrdetectionofthehhaitandemdnarepeatinpreandpostpatentbrugiamalayiinfectionsastudyinindonesiantransmigrants
AT hoeraufachim realtimepcrdetectionofthehhaitandemdnarepeatinpreandpostpatentbrugiamalayiinfectionsastudyinindonesiantransmigrants