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通过针对一个基因序列的实时聚合酶链反应(PCR)对公牛进行准确快速的鉴定。

Accurate and fast identification of in bulls by real-time PCR targeting a gene sequence.

作者信息

Delpiazzo Rafael, Barcellos Maila, Barros Sofía, Betancor Laura, Fraga Martín, Gil Jorge, Iraola Gregorio, Morsella Claudia, Paolicchi Fernando, Pérez Ruben, Riet-Correa Franklin, Sanguinetti Margarita, Silva Alfonso, da Silva Silveira Caroline, Calleros Lucía

机构信息

Departamento de Salud de los Sistemas Pecuarios, Facultad de Veterinaria, Universidad de la República Oriental del Uruguay, Estación Experimental "Dr. Mario A. Cassinoni", Ruta 3 Km. 363, Paysandú, Uruguay.

Sección Genética Evolutiva, Facultad de Ciencias, Universidad de la República Oriental del Uruguay, Iguá 4225, Montevideo, Uruguay.

出版信息

Vet Anim Sci. 2020 Dec 24;11:100163. doi: 10.1016/j.vas.2020.100163. eCollection 2021 Mar.

Abstract

is an important animal pathogen that causes infectious infertility, embryonic mortality and abortions in cattle and sheep flocks. There are two recognized subspecies related with reproductive disorders in livestock: subsp. (Cff) and subsp. (Cfv). Rapid and reliable detection of this pathogenic species in bulls is of upmost importance for disease control in dairy and beef herds as they are asymptomatic carriers. The aim of the present work was to assess the performance a real-time PCR (qPCR) method for the diagnosis of in samples from bulls, comparing it with culture and isolation methods. 520 preputial samples were both cultured in Skirrow's medium and analyzed by qPCR. The estimated sensitivity of qPCR was 90.9% (95% CI, 69.4%-100%), and the specificity was 99.4% (95% CI, 98.6% - 100%). The proportion of positive individuals was 2.1% by isolation and 2.5% by qPCR. Isolates were identified by biochemical tests as Cfv ( = 9) and Cff ( = 2). Our findings support the use of qPCR for fast and accurate detection of directly from field samples of preputial smegma of bulls. The qPCR method showed to be suitable for massive screenings because it can be performed in pooled samples without losing accuracy and sensitivity.

摘要

是一种重要的动物病原体,可导致牛羊群感染性不育、胚胎死亡和流产。有两个公认的与家畜生殖障碍相关的亚种:亚种(Cff)和亚种(Cfv)。由于公牛是无症状携带者,因此在公牛中快速可靠地检测这种致病物种对于奶牛和肉牛群的疾病控制至关重要。本研究的目的是评估实时荧光定量聚合酶链反应(qPCR)方法在诊断公牛样本中的性能,并将其与培养和分离方法进行比较。520份包皮样本分别在Skirrow培养基中培养并通过qPCR进行分析。qPCR的估计灵敏度为90.9%(95%CI,69.4%-100%),特异性为99.4%(95%CI,98.6%-100%)。通过分离法检测到的阳性个体比例为2.1%,通过qPCR检测到的阳性个体比例为2.5%。通过生化试验将分离株鉴定为Cfv(=9)和Cff(=2)。我们的研究结果支持使用qPCR直接从公牛包皮垢的现场样本中快速准确地检测。qPCR方法适用于大规模筛查,因为它可以在混合样本中进行,而不会损失准确性和灵敏度。

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