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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.<\/jats:p>\n              <\/jats:sec>\n              <jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>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.<\/jats:p>\n              <\/jats:sec>\n              <jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>A multiplex polymerase chain reaction (PCR) assay was developed based on mitochondrial DNA (<jats:italic>mt<\/jats:italic>DNA) that accurately and simultaneously identify four cestode species in one reaction using specific fragment sizes of 592, 385, 283, and 190\u2009bp for <jats:italic>T. hydatigena<\/jats:italic>, <jats:italic>T. multiceps<\/jats:italic>, <jats:italic>T. pisiformis<\/jats:italic>, and <jats:italic>D. caninum<\/jats:italic>, respectively. The lowest DNA concentration detected was 1\u2009ng for <jats:italic>T. hydatigena<\/jats:italic>, <jats:italic>T. multiceps<\/jats:italic> and <jats:italic>T. pisiformis<\/jats:italic>, and 0.1\u2009ng for <jats:italic>D. caninum<\/jats:italic> in a 25\u2009\u03bcl reaction system. This assay offers high potential for the rapid detection of these four tapeworms in host feces simultaneously.<\/jats:p>\n              <\/jats:sec>\n              <jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>This study provides an efficient tool for the simultaneous detection of <jats:italic>T. hydatigena<\/jats:italic>, <jats:italic>T. multiceps<\/jats:italic>, <jats:italic>T. pisiformis<\/jats:italic>, and <jats:italic>D. caninum<\/jats:italic>. The assay will be potentially useful in epidemiological studies, diagnosis, and treatment of these four cestodes infections during prevention and control program.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1186\/s12879-019-4512-3","type":"journal-article","created":{"date-parts":[[2019,10,16]],"date-time":"2019-10-16T12:07:53Z","timestamp":1571227673000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections"],"prefix":"10.1186","volume":"19","author":[{"given":"Guo-Qiang","family":"Zhu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John Asekhaen","family":"Ohiolei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan-Tao","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wen-Hui","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nian-Zhang","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bao-Quan","family":"Fu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong-Bin","family":"Yan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wan-Zhong","family":"Jia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2019,10,16]]},"reference":[{"issue":"8","key":"4512_CR1","doi-asserted-by":"publisher","first-page":"859","DOI":"10.1016\/S1286-4579(02)01606-4","volume":"4","author":"EP Hoberg","year":"2002","unstructured":"Hoberg EP. 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The study was approved by the Animal Ethics Committee of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences (No. LVRIAEC2012\u2013007).","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare that they have no competing interests.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"854"}}