Development and application of a multiplex TaqMan® real-time qPCR assay for the simultaneous detection of Anaplasma marginale and Theileria annulata and molecular characterization of Anaplasma marginale from cattle in Western Cuba
| العنوان: | Development and application of a multiplex TaqMan® real-time qPCR assay for the simultaneous detection of Anaplasma marginale and Theileria annulata and molecular characterization of Anaplasma marginale from cattle in Western Cuba |
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| المؤلفون: | Adrian Alberto Díaz-Sánchez, Dasiel Obregón Alvarez, Belkis Corona-González, Alejandro Cabezas-Cruz, Regina Hofmann-Lehmann, Marina L. Meli, Osvaldo Fonseca-Rodríguez |
| المساهمون: | Centro Nacional de Sanidad Agropecuaria (CENSA), Universität Zürich [Zürich] = University of Zurich (UZH), University of Guelph, University of São Paulo (USP), Umeå University, Biologie moléculaire et immunologie parasitaires et fongiques (BIPAR), École nationale vétérinaire d'Alfort (ENVA)-Laboratoire de santé animale, sites de Maisons-Alfort et de Dozulé, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Swiss Government Excellence Scholarship - Federal Commission for Scholarships for Foreign Students (FCS) 2016.0828, École nationale vétérinaire - Alfort (ENVA)-Laboratoire de santé animale, sites de Maisons-Alfort et de Dozulé, University of Zurich, Corona-González, Belkis |
| المصدر: | Ticks and Tick-Borne Diseases Ticks and Tick-Borne Diseases, 2019, 11 (2), ⟨10.1016/j.ttbdis.2019.101356⟩ Ticks and Tick-Borne Diseases, 2020, 11 (2), pp.101356. ⟨10.1016/j.ttbdis.2019.101356⟩ |
| بيانات النشر: | Elsevier BV, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | 0301 basic medicine, Anaplasmosis, 1109 Insect Science, MSP1a, [SDV]Life Sciences [q-bio], 030231 tropical medicine, 2405 Parasitology, Cattle Diseases, 610 Medicine & health, Biology, Real-Time Polymerase Chain Reaction, Microbiology, 03 medical and health sciences, 0302 clinical medicine, Tandem repeat, Genotype, medicine, TaqMan, Animals, Multiplex, 11434 Center for Clinical Studies, 2. Zero hunger, MSP4, 2404 Microbiology, Cuba, Reproducibility of Results, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, 2725 Infectious Diseases, 16S ribosomal RNA, medicine.disease, Theileria annulata, Virology, Theileriasis, Anaplasma marginale, 030104 developmental biology, Infectious Diseases, Multiplex TaqMan®qPCR, Insect Science, 11404 Department of Clinical Diagnostics and Services, Microsatellite, Cattle, Parasitology, Multiplex Polymerase Chain Reaction, Nested polymerase chain reaction |
| الوصف: | International audience; Anaplasmosis and theileriosis are considered the most important tick-borne diseases for livestock production worldwide, causing significant economic losses in tropical and subtropical regions. The present study was aimed to develop a multiplex TaqMan® qPCR assay to simultaneously detect Anaplasma marginale and Theileria annulata and to applied it to investigate naturally infected cattle in Cuba. The assay was highly specific, sensible, and efficient; it was more sensitive than a well-established nested PCR and detected 1 DNA copy of each target. Consistent repeatability and reproducibility within and between multiplex qPCR runs was shown. A total of 223 blood samples collected in western Cuba were analyzed for haemoparasites infection in cattle. The multiplex qPCR assay detected A. marginale in 213 samples (95.5%; CI: 95%; 91.9%-97.5%), but all samples were negative for T. annulata. Additionally, the genetic diversity of A. marginale was assessed using 16S rRNA, MSP1a and MSP4 nucleotide and protein sequences. The MSP1a tandem repeats ranged from three to five, and twelve different MSP1a tandem repeats of A. marginale were found, which presented genotypes C, E, and G in the 5ʹUTR microsatellite region. Phylogenetic analysis using the msp4 gene showed that Cuban strains were closely related to others previously reported in Mexico, Brazil and Asian countries. The multiplex qPCR described here proved to be a rapid, specific and cost-effective mean for the simultaneous detection of A. marginale and T. annulata. Further epidemiological studies using this assay will improve the surveillance of the associated diseases in regions where they are endemic. |
| وصف الملف: | 1-s2.0-S1877959X19302596-main.pdf - application/pdf |
| تدمد: | 1877-959X |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::1d24eda3356033747760ee0f2d3dee90Test https://doi.org/10.1016/j.ttbdis.2019.101356Test |
| حقوق: | RESTRICTED |
| رقم الانضمام: | edsair.doi.dedup.....1d24eda3356033747760ee0f2d3dee90 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 1877959X |
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