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High resolution melting analysis of the 18S rRNA gene for the rapid diagnosis of bovine babesiosis

BACKGROUND: Bovine babesiosis is caused by protozoan parasites of the genus Babesia and presents a wide spectrum of clinical manifestations. Disease severity depends on the type of Babesia species infection. Generally, B. bovis and B. bigemina are considered as the causative agents of bovine babesio...

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Autores principales: Wang, Jinming, Liu, Aihong, Zhang, Shangdi, Gao, Shandian, Rashid, Muhammad, Li, Youquan, Liu, Junlong, Ma, Quanying, Li, Zhi, Liu, Zhijie, Luo, Jianxun, Guan, Guiquan, Yin, Hong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6833191/
https://www.ncbi.nlm.nih.gov/pubmed/31694676
http://dx.doi.org/10.1186/s13071-019-3781-4
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author Wang, Jinming
Liu, Aihong
Zhang, Shangdi
Gao, Shandian
Rashid, Muhammad
Li, Youquan
Liu, Junlong
Ma, Quanying
Li, Zhi
Liu, Zhijie
Luo, Jianxun
Guan, Guiquan
Yin, Hong
author_facet Wang, Jinming
Liu, Aihong
Zhang, Shangdi
Gao, Shandian
Rashid, Muhammad
Li, Youquan
Liu, Junlong
Ma, Quanying
Li, Zhi
Liu, Zhijie
Luo, Jianxun
Guan, Guiquan
Yin, Hong
author_sort Wang, Jinming
collection PubMed
description BACKGROUND: Bovine babesiosis is caused by protozoan parasites of the genus Babesia and presents a wide spectrum of clinical manifestations. Disease severity depends on the type of Babesia species infection. Generally, B. bovis and B. bigemina are considered as the causative agents of bovine babesiosis; in addition, Babesia ovata and B. major are a group of benign bovine piroplasms. Therefore, species identification is important for diagnosis, epidemiological investigations and follow-up management. METHODS: Real-time PCR combined with high resolution melting (RT-PCR-HRM) analysis was used to detect and discriminate four Babesia species infective to cattle, including Babesia bovis, B. bigemina, B. major and B. ovata. The melting profiles and melting temperatures (Tm) of the amplicon targeting 18S rRNA revealed differences that can discriminate the four Babesia spp. Sensitivity and specificity of the analytical method were evaluated using 50 blood samples collected from experimentally infected cattle and 240 blood samples from areas where bovine babesiosis is an issue. RESULTS: RT-PCR-HRM analysis allowed to detect and discriminate four Babesia spp. (B. bovis, B. bigemina, B. major and B. ovata), which were responsible for bovine babesiosis in China. The protocol was validated with DNA samples from experimentally infected cattle and field infection in cattle. CONCLUSIONS: Our results indicate that RT-PCR-HRM is a fast and robust tool for the simultaneous detection and discrimination of four Babesia species that are responsible for bovine babesiosis in China. This approach is applicable for both field and experimental samples, thus it could be useful in epidemiological investigations and diagnoses of bovine babesiosis.
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spelling pubmed-68331912019-11-08 High resolution melting analysis of the 18S rRNA gene for the rapid diagnosis of bovine babesiosis Wang, Jinming Liu, Aihong Zhang, Shangdi Gao, Shandian Rashid, Muhammad Li, Youquan Liu, Junlong Ma, Quanying Li, Zhi Liu, Zhijie Luo, Jianxun Guan, Guiquan Yin, Hong Parasit Vectors Research BACKGROUND: Bovine babesiosis is caused by protozoan parasites of the genus Babesia and presents a wide spectrum of clinical manifestations. Disease severity depends on the type of Babesia species infection. Generally, B. bovis and B. bigemina are considered as the causative agents of bovine babesiosis; in addition, Babesia ovata and B. major are a group of benign bovine piroplasms. Therefore, species identification is important for diagnosis, epidemiological investigations and follow-up management. METHODS: Real-time PCR combined with high resolution melting (RT-PCR-HRM) analysis was used to detect and discriminate four Babesia species infective to cattle, including Babesia bovis, B. bigemina, B. major and B. ovata. The melting profiles and melting temperatures (Tm) of the amplicon targeting 18S rRNA revealed differences that can discriminate the four Babesia spp. Sensitivity and specificity of the analytical method were evaluated using 50 blood samples collected from experimentally infected cattle and 240 blood samples from areas where bovine babesiosis is an issue. RESULTS: RT-PCR-HRM analysis allowed to detect and discriminate four Babesia spp. (B. bovis, B. bigemina, B. major and B. ovata), which were responsible for bovine babesiosis in China. The protocol was validated with DNA samples from experimentally infected cattle and field infection in cattle. CONCLUSIONS: Our results indicate that RT-PCR-HRM is a fast and robust tool for the simultaneous detection and discrimination of four Babesia species that are responsible for bovine babesiosis in China. This approach is applicable for both field and experimental samples, thus it could be useful in epidemiological investigations and diagnoses of bovine babesiosis. BioMed Central 2019-11-06 /pmc/articles/PMC6833191/ /pubmed/31694676 http://dx.doi.org/10.1186/s13071-019-3781-4 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
Wang, Jinming
Liu, Aihong
Zhang, Shangdi
Gao, Shandian
Rashid, Muhammad
Li, Youquan
Liu, Junlong
Ma, Quanying
Li, Zhi
Liu, Zhijie
Luo, Jianxun
Guan, Guiquan
Yin, Hong
High resolution melting analysis of the 18S rRNA gene for the rapid diagnosis of bovine babesiosis
title High resolution melting analysis of the 18S rRNA gene for the rapid diagnosis of bovine babesiosis
title_full High resolution melting analysis of the 18S rRNA gene for the rapid diagnosis of bovine babesiosis
title_fullStr High resolution melting analysis of the 18S rRNA gene for the rapid diagnosis of bovine babesiosis
title_full_unstemmed High resolution melting analysis of the 18S rRNA gene for the rapid diagnosis of bovine babesiosis
title_short High resolution melting analysis of the 18S rRNA gene for the rapid diagnosis of bovine babesiosis
title_sort high resolution melting analysis of the 18s rrna gene for the rapid diagnosis of bovine babesiosis
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6833191/
https://www.ncbi.nlm.nih.gov/pubmed/31694676
http://dx.doi.org/10.1186/s13071-019-3781-4
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