Cargando…

A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections

BACKGROUND: Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum are four common large and medium-sized tapeworms parasitizing the small intestine of dogs and other canids. These parasites cause serious impact on the health and development of livestock. However, there are, so far,...

Descripción completa

Detalles Bibliográficos
Autores principales: Zhu, Guo-Qiang, Li, Li, Ohiolei, John Asekhaen, Wu, Yan-Tao, Li, Wen-Hui, Zhang, Nian-Zhang, Fu, Bao-Quan, Yan, Hong-Bin, Jia, Wan-Zhong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6796438/
https://www.ncbi.nlm.nih.gov/pubmed/31619188
http://dx.doi.org/10.1186/s12879-019-4512-3
_version_ 1783459589997461504
author Zhu, Guo-Qiang
Li, Li
Ohiolei, John Asekhaen
Wu, Yan-Tao
Li, Wen-Hui
Zhang, Nian-Zhang
Fu, Bao-Quan
Yan, Hong-Bin
Jia, Wan-Zhong
author_facet Zhu, Guo-Qiang
Li, Li
Ohiolei, John Asekhaen
Wu, Yan-Tao
Li, Wen-Hui
Zhang, Nian-Zhang
Fu, Bao-Quan
Yan, Hong-Bin
Jia, Wan-Zhong
author_sort Zhu, Guo-Qiang
collection PubMed
description BACKGROUND: Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum are four common large and medium-sized tapeworms parasitizing the small intestine of dogs and other canids. These parasites cause serious impact on the health and development of livestock. However, there are, so far, no commercially available molecular diagnostic kits capable of simultaneously detecting all four parasites in dogs. The aim of the study was therefore to develop a multiplex PCR assay that will accurately detect all four cestode infections in one reaction. METHODS: Specific primers for a multiplex PCR were designed based on corresponding mitochondrial genome sequences, and its detection limit was assessed by serial dilutions of the genomic DNAs of tapeworms examined. Furthermore, field samples of dog feces were tested using the developed assay. RESULTS: A multiplex polymerase chain reaction (PCR) assay was developed based on mitochondrial DNA (mtDNA) that accurately and simultaneously identify four cestode species in one reaction using specific fragment sizes of 592, 385, 283, and 190 bp for T. hydatigena, T. multiceps, T. pisiformis, and D. caninum, respectively. The lowest DNA concentration detected was 1 ng for T. hydatigena, T. multiceps and T. pisiformis, and 0.1 ng for D. caninum in a 25 μl reaction system. This assay offers high potential for the rapid detection of these four tapeworms in host feces simultaneously. CONCLUSIONS: This study provides an efficient tool for the simultaneous detection of T. hydatigena, T. multiceps, T. pisiformis, and D. caninum. The assay will be potentially useful in epidemiological studies, diagnosis, and treatment of these four cestodes infections during prevention and control program.
format Online
Article
Text
id pubmed-6796438
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-67964382019-10-21 A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections Zhu, Guo-Qiang Li, Li Ohiolei, John Asekhaen Wu, Yan-Tao Li, Wen-Hui Zhang, Nian-Zhang Fu, Bao-Quan Yan, Hong-Bin Jia, Wan-Zhong BMC Infect Dis Research Article BACKGROUND: Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum are four common large and medium-sized tapeworms parasitizing the small intestine of dogs and other canids. These parasites cause serious impact on the health and development of livestock. However, there are, so far, no commercially available molecular diagnostic kits capable of simultaneously detecting all four parasites in dogs. The aim of the study was therefore to develop a multiplex PCR assay that will accurately detect all four cestode infections in one reaction. METHODS: Specific primers for a multiplex PCR were designed based on corresponding mitochondrial genome sequences, and its detection limit was assessed by serial dilutions of the genomic DNAs of tapeworms examined. Furthermore, field samples of dog feces were tested using the developed assay. RESULTS: A multiplex polymerase chain reaction (PCR) assay was developed based on mitochondrial DNA (mtDNA) that accurately and simultaneously identify four cestode species in one reaction using specific fragment sizes of 592, 385, 283, and 190 bp for T. hydatigena, T. multiceps, T. pisiformis, and D. caninum, respectively. The lowest DNA concentration detected was 1 ng for T. hydatigena, T. multiceps and T. pisiformis, and 0.1 ng for D. caninum in a 25 μl reaction system. This assay offers high potential for the rapid detection of these four tapeworms in host feces simultaneously. CONCLUSIONS: This study provides an efficient tool for the simultaneous detection of T. hydatigena, T. multiceps, T. pisiformis, and D. caninum. The assay will be potentially useful in epidemiological studies, diagnosis, and treatment of these four cestodes infections during prevention and control program. BioMed Central 2019-10-16 /pmc/articles/PMC6796438/ /pubmed/31619188 http://dx.doi.org/10.1186/s12879-019-4512-3 Text en © The Author(s). 2019 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 Article
Zhu, Guo-Qiang
Li, Li
Ohiolei, John Asekhaen
Wu, Yan-Tao
Li, Wen-Hui
Zhang, Nian-Zhang
Fu, Bao-Quan
Yan, Hong-Bin
Jia, Wan-Zhong
A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections
title A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections
title_full A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections
title_fullStr A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections
title_full_unstemmed A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections
title_short A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections
title_sort multiplex pcr assay for the simultaneous detection of taenia hydatigena, t. multiceps, t. pisiformis, and dipylidium caninum infections
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6796438/
https://www.ncbi.nlm.nih.gov/pubmed/31619188
http://dx.doi.org/10.1186/s12879-019-4512-3
work_keys_str_mv AT zhuguoqiang amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT lili amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT ohioleijohnasekhaen amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT wuyantao amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT liwenhui amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT zhangnianzhang amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT fubaoquan amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT yanhongbin amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT jiawanzhong amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT zhuguoqiang multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT lili multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT ohioleijohnasekhaen multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT wuyantao multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT liwenhui multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT zhangnianzhang multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT fubaoquan multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT yanhongbin multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections
AT jiawanzhong multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections