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HDP2: a ribosomal DNA (NTS-ETS) sequence as a target for species-specific molecular diagnosis of intestinal taeniasis in humans

BACKGROUND: Taenia solium, T. asiatica and T. saginata tapeworms cause human taeniasis and are the origin of porcine and bovine cysticercosis. Furthermore, T. solium eggs can cause human cysticercosis, with neurocysticercosis being the most serious form of the disease. These helminth infections are...

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Autores principales: Flores, María D., Gonzalez, Luis M., Hurtado, Carolina, Motta, Yamileth Monje, Domínguez-Hidalgo, Cristina, Merino, Francisco Jesús, Perteguer, María J., Gárate, Teresa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6389257/
https://www.ncbi.nlm.nih.gov/pubmed/29486767
http://dx.doi.org/10.1186/s13071-018-2646-6
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author Flores, María D.
Gonzalez, Luis M.
Hurtado, Carolina
Motta, Yamileth Monje
Domínguez-Hidalgo, Cristina
Merino, Francisco Jesús
Perteguer, María J.
Gárate, Teresa
author_facet Flores, María D.
Gonzalez, Luis M.
Hurtado, Carolina
Motta, Yamileth Monje
Domínguez-Hidalgo, Cristina
Merino, Francisco Jesús
Perteguer, María J.
Gárate, Teresa
author_sort Flores, María D.
collection PubMed
description BACKGROUND: Taenia solium, T. asiatica and T. saginata tapeworms cause human taeniasis and are the origin of porcine and bovine cysticercosis. Furthermore, T. solium eggs can cause human cysticercosis, with neurocysticercosis being the most serious form of the disease. These helminth infections are neglected tropical diseases and are endemic in several countries in the Americas, Asia and Africa. As a result of globalization, migration in particular, the infections have been extending to non-endemic territories. Species-specific diagnosis of taeniasis is subject to drawbacks that could be resolved using molecular approaches. In the present study, conventional and real-time amplification protocols (cPCR and qPCR) based on the T. saginata HDP2 sequence were applied in the differential diagnosis of taeniasis (T. saginata, T. solium) in both fecal samples and proglottids expelled by patients. The HDP2 homolog in T. solium was cloned and characterized. RESULTS: Semi-nested cPCR and qPCR (Sn-HDP2 cPCR and Sn-HDP2 qPCR) amplified T. saginata and T. solium DNA, with an analytical sensitivity of 40 and 400 fg, respectively, and identically in both protocols. Eighteen taeniasis patients were diagnosed directly with T. saginata or T. solium, either from proglottids or fecal samples with/without eggs (detected using microscopy), based on the optimized Sn-HDP2 qPCR. After cloning, the T. solium HDP2 homolog sequence was confirmed to be a ribosomal sequence. The HDP2 fragment corresponded to a non-transcribed sequence/external transcribed repeat (NTS/ETS) of ribosomal DNA. Compared with the T. saginata HDP2 homolog, the T solium HDP2 sequence lacked the first 900 nt at the 5′ end and showed nucleotide substitutions and small deletions. CONCLUSIONS: Sn-HDP2 cPCR and Sn-HDP2 qPCR were set up for the diagnosis of human taeniasis, using proglottids and fecal samples from affected patients. The new Sn-HDP2 qPCR protocol was the best option, as it directly differentiated T. saginata from T. solium. The diagnosis of an imported T. solium-taeniasis case and nine European T. saginata cases was relevant. Finally, the cloning and sequencing of the T. solium HDP2 fragment confirmed that HDP2 was part of a ribosomal unit. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13071-018-2646-6) contains supplementary material, which is available to authorized users.
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spelling pubmed-63892572019-03-19 HDP2: a ribosomal DNA (NTS-ETS) sequence as a target for species-specific molecular diagnosis of intestinal taeniasis in humans Flores, María D. Gonzalez, Luis M. Hurtado, Carolina Motta, Yamileth Monje Domínguez-Hidalgo, Cristina Merino, Francisco Jesús Perteguer, María J. Gárate, Teresa Parasit Vectors Research BACKGROUND: Taenia solium, T. asiatica and T. saginata tapeworms cause human taeniasis and are the origin of porcine and bovine cysticercosis. Furthermore, T. solium eggs can cause human cysticercosis, with neurocysticercosis being the most serious form of the disease. These helminth infections are neglected tropical diseases and are endemic in several countries in the Americas, Asia and Africa. As a result of globalization, migration in particular, the infections have been extending to non-endemic territories. Species-specific diagnosis of taeniasis is subject to drawbacks that could be resolved using molecular approaches. In the present study, conventional and real-time amplification protocols (cPCR and qPCR) based on the T. saginata HDP2 sequence were applied in the differential diagnosis of taeniasis (T. saginata, T. solium) in both fecal samples and proglottids expelled by patients. The HDP2 homolog in T. solium was cloned and characterized. RESULTS: Semi-nested cPCR and qPCR (Sn-HDP2 cPCR and Sn-HDP2 qPCR) amplified T. saginata and T. solium DNA, with an analytical sensitivity of 40 and 400 fg, respectively, and identically in both protocols. Eighteen taeniasis patients were diagnosed directly with T. saginata or T. solium, either from proglottids or fecal samples with/without eggs (detected using microscopy), based on the optimized Sn-HDP2 qPCR. After cloning, the T. solium HDP2 homolog sequence was confirmed to be a ribosomal sequence. The HDP2 fragment corresponded to a non-transcribed sequence/external transcribed repeat (NTS/ETS) of ribosomal DNA. Compared with the T. saginata HDP2 homolog, the T solium HDP2 sequence lacked the first 900 nt at the 5′ end and showed nucleotide substitutions and small deletions. CONCLUSIONS: Sn-HDP2 cPCR and Sn-HDP2 qPCR were set up for the diagnosis of human taeniasis, using proglottids and fecal samples from affected patients. The new Sn-HDP2 qPCR protocol was the best option, as it directly differentiated T. saginata from T. solium. The diagnosis of an imported T. solium-taeniasis case and nine European T. saginata cases was relevant. Finally, the cloning and sequencing of the T. solium HDP2 fragment confirmed that HDP2 was part of a ribosomal unit. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13071-018-2646-6) contains supplementary material, which is available to authorized users. BioMed Central 2018-02-27 /pmc/articles/PMC6389257/ /pubmed/29486767 http://dx.doi.org/10.1186/s13071-018-2646-6 Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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
Flores, María D.
Gonzalez, Luis M.
Hurtado, Carolina
Motta, Yamileth Monje
Domínguez-Hidalgo, Cristina
Merino, Francisco Jesús
Perteguer, María J.
Gárate, Teresa
HDP2: a ribosomal DNA (NTS-ETS) sequence as a target for species-specific molecular diagnosis of intestinal taeniasis in humans
title HDP2: a ribosomal DNA (NTS-ETS) sequence as a target for species-specific molecular diagnosis of intestinal taeniasis in humans
title_full HDP2: a ribosomal DNA (NTS-ETS) sequence as a target for species-specific molecular diagnosis of intestinal taeniasis in humans
title_fullStr HDP2: a ribosomal DNA (NTS-ETS) sequence as a target for species-specific molecular diagnosis of intestinal taeniasis in humans
title_full_unstemmed HDP2: a ribosomal DNA (NTS-ETS) sequence as a target for species-specific molecular diagnosis of intestinal taeniasis in humans
title_short HDP2: a ribosomal DNA (NTS-ETS) sequence as a target for species-specific molecular diagnosis of intestinal taeniasis in humans
title_sort hdp2: a ribosomal dna (nts-ets) sequence as a target for species-specific molecular diagnosis of intestinal taeniasis in humans
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6389257/
https://www.ncbi.nlm.nih.gov/pubmed/29486767
http://dx.doi.org/10.1186/s13071-018-2646-6
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