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Development of a real-time PCR method for rapid diagnosis of canine babesiosis and anaplasmosis
BACKGROUND: Canine babesiosis and anaplasmosis, caused by Babesia canis and Anaplasma phagocytophilum, respectively, are significant tick-borne diseases in Baltic countries. Both diseases can be diagnosed on the basis of clinicopathological findings, by direct pathogen detection in blood smears or b...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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BioMed Central
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8139040/ https://www.ncbi.nlm.nih.gov/pubmed/34016173 http://dx.doi.org/10.1186/s13071-021-04756-9 |
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author | Kivrane, Agnija Namina, Agne Seleznova, Maija Akopjana, Sarmite Capligina, Valentina Ranka, Renate |
author_facet | Kivrane, Agnija Namina, Agne Seleznova, Maija Akopjana, Sarmite Capligina, Valentina Ranka, Renate |
author_sort | Kivrane, Agnija |
collection | PubMed |
description | BACKGROUND: Canine babesiosis and anaplasmosis, caused by Babesia canis and Anaplasma phagocytophilum, respectively, are significant tick-borne diseases in Baltic countries. Both diseases can be diagnosed on the basis of clinicopathological findings, by direct pathogen detection in blood smears or by indirect pathogen detection; however, because of high selectivity and specificity, molecular methods may be advantageous. The goal of this study was to develop a duplex real-time polymerase chain reaction (RT-PCR) method for the detection of B. canis and A. phagocytophilum in canine clinical samples. METHODS: Sequence-based polymorphism analysis of genes encoding B. canis-specific merozoite surface protein Bc28.1 (Bc28.1) and A. phagocytophilum malate dehydrogenase (mdh) was performed on pathogen isolates present in Latvian domestic dogs. The obtained results were used to design a species-specific duplex RT-PCR assay. RESULTS: The presence of three B. canis Bc28.1 gene sequence types was revealed in canine samples with a nonuniform geographical distribution, and two types of A. phagocytophilum mdh genes were detected. The novel duplex RT-PCR assay provided correct classification of samples positive and negative for B. canis and A. phagocytophilum. The analytical sensitivity of this assay was ten gene copies/ reaction for both pathogens. CONCLUSIONS: A novel duplex RT-PCR molecular method was developed for the detection of B. canis and A. phagocytophilum in canine clinical samples. Sequence variability of Bc28.1 and mdh genes indicated the genetic variability of B. canis and A. phagocytophilum isolates occurring in Latvian domestic dogs. GRAPHIC ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13071-021-04756-9. |
format | Online Article Text |
id | pubmed-8139040 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-81390402021-05-21 Development of a real-time PCR method for rapid diagnosis of canine babesiosis and anaplasmosis Kivrane, Agnija Namina, Agne Seleznova, Maija Akopjana, Sarmite Capligina, Valentina Ranka, Renate Parasit Vectors Research BACKGROUND: Canine babesiosis and anaplasmosis, caused by Babesia canis and Anaplasma phagocytophilum, respectively, are significant tick-borne diseases in Baltic countries. Both diseases can be diagnosed on the basis of clinicopathological findings, by direct pathogen detection in blood smears or by indirect pathogen detection; however, because of high selectivity and specificity, molecular methods may be advantageous. The goal of this study was to develop a duplex real-time polymerase chain reaction (RT-PCR) method for the detection of B. canis and A. phagocytophilum in canine clinical samples. METHODS: Sequence-based polymorphism analysis of genes encoding B. canis-specific merozoite surface protein Bc28.1 (Bc28.1) and A. phagocytophilum malate dehydrogenase (mdh) was performed on pathogen isolates present in Latvian domestic dogs. The obtained results were used to design a species-specific duplex RT-PCR assay. RESULTS: The presence of three B. canis Bc28.1 gene sequence types was revealed in canine samples with a nonuniform geographical distribution, and two types of A. phagocytophilum mdh genes were detected. The novel duplex RT-PCR assay provided correct classification of samples positive and negative for B. canis and A. phagocytophilum. The analytical sensitivity of this assay was ten gene copies/ reaction for both pathogens. CONCLUSIONS: A novel duplex RT-PCR molecular method was developed for the detection of B. canis and A. phagocytophilum in canine clinical samples. Sequence variability of Bc28.1 and mdh genes indicated the genetic variability of B. canis and A. phagocytophilum isolates occurring in Latvian domestic dogs. GRAPHIC ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13071-021-04756-9. BioMed Central 2021-05-20 /pmc/articles/PMC8139040/ /pubmed/34016173 http://dx.doi.org/10.1186/s13071-021-04756-9 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Kivrane, Agnija Namina, Agne Seleznova, Maija Akopjana, Sarmite Capligina, Valentina Ranka, Renate Development of a real-time PCR method for rapid diagnosis of canine babesiosis and anaplasmosis |
title | Development of a real-time PCR method for rapid diagnosis of canine babesiosis and anaplasmosis |
title_full | Development of a real-time PCR method for rapid diagnosis of canine babesiosis and anaplasmosis |
title_fullStr | Development of a real-time PCR method for rapid diagnosis of canine babesiosis and anaplasmosis |
title_full_unstemmed | Development of a real-time PCR method for rapid diagnosis of canine babesiosis and anaplasmosis |
title_short | Development of a real-time PCR method for rapid diagnosis of canine babesiosis and anaplasmosis |
title_sort | development of a real-time pcr method for rapid diagnosis of canine babesiosis and anaplasmosis |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8139040/ https://www.ncbi.nlm.nih.gov/pubmed/34016173 http://dx.doi.org/10.1186/s13071-021-04756-9 |
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