دورية أكاديمية
New sensitive real-time PCR targeting p28 gene for detection of Ehrlichia canis in blood samples from dogs
العنوان: | New sensitive real-time PCR targeting p28 gene for detection of Ehrlichia canis in blood samples from dogs |
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المؤلفون: | Patrícia Gonzaga Paulino, Tays Araujo Camilo, Miguel Angelo Leite Mota Junior, Nathália Alves de Senne, Olga Lucia Herrán Ramirez, Renata Lins da Costa, Carlos Luiz Massard, Huarrisson Azevedo Santos |
المصدر: | Ciência Rural, Vol 51, Iss 12 (2021) |
بيانات النشر: | Universidade Federal de Santa Maria, 2021. |
سنة النشر: | 2021 |
المجموعة: | LCC:Agriculture LCC:Agriculture (General) |
مصطلحات موضوعية: | canine monocytic ehrlichiosis, molecular detection, diagnosis, hemoparasite, Agriculture, Agriculture (General), S1-972 |
الوصف: | ABSTRACT: This study aims to describe a new detection method of a quantitative real-time polymerase chain reaction (qPCR) targeting the 28 kDa outer membrane protein gene (p28) as well as to compare this method with a conventional PCR (cPCR), which targets the same gene, in order to evaluate the performance of the technique designed in this study in detecting Ehrlichia canis (E. canis). Optimum oligonucleotides concentrations were reached, and the analytical sensitivity and specificity of the qPCR were performed. A total of 218 dogs’ whole blood samples were conventionally collected for this study. The DNA was extracted from each sample. Subsequently, the samples were tested by an established cPCR and the new qPCR to compare each technique’s performances. This new qPCR method for the molecular detection of E. canis presented a detection limit of ten copies of the fragment and was considered specific for E. canis according to analytical specificity analyses performed in vitro and in silico. The standard curve revealed 100% efficiency and a coefficient of determination (R2) equivalent to 99.8%. Among the samples examined by qPCR, 24.31% were considered positive, significantly greater than those detected by cPCR (15.13%). The qPCR technique reached a higher sensitivity than the cPCR when targeting the p28 gene in detecting E. canis. The qPCR standardized in this study is an efficient method for confirming canine monocytic ehrlichiosis (CME) diagnosis and might provide the parasitemia monitoring during the disease treatment. |
نوع الوثيقة: | article |
وصف الملف: | electronic resource |
اللغة: | English Portuguese |
تدمد: | 1678-4596 0103-8478 |
العلاقة: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782021001200454&tlng=enTest; https://doaj.org/toc/1678-4596Test |
DOI: | 10.1590/0103-8478cr20200891 |
الوصول الحر: | https://doaj.org/article/e04d1131ad18494199f1645b484c1b6cTest |
رقم الانضمام: | edsdoj.04d1131ad18494199f1645b484c1b6c |
قاعدة البيانات: | Directory of Open Access Journals |
تدمد: | 16784596 01038478 |
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DOI: | 10.1590/0103-8478cr20200891 |