分子检验在诊断布鲁氏菌引起的绵羊布鲁氏菌病中的可靠性

Mladen Zelenović, Darko Marinković, Nataša Stević, Slavoljub Stanojević, Milan Aničić, Vesna Milićević, Olivera Valčić, Sonja Radojičić
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摘要

布鲁氏菌(Brucella ovis)会感染绵羊,导致临床或亚临床疾病,其特征是公羊生殖器病变和繁殖力下降,母羊胎盘炎和流产,以及羔羊围产期死亡率增加。作为这项研究的一部分,在贝尔格莱德(塞尔维亚)附近的一个养羊场进行了分析。在血清学检测中,进行了间接 ELISA 检测。共分析了 94 份血清,其中 33 份来自公羊,61 份来自母羊。结果显示,公羊和母羊的阳性结果分别为 23 例(69.7%)和 2 例(3.3%),总患病率为 25.4%,置信区间为 95%。对 19 只血清学检测呈阳性的公羊和 2 只血清学检测呈阳性的母羊进行了布鲁斯-梯状多重 PCR 和实时 PCR 检测。结果表明,与传统 PCR 相比,Real time PCR 在诊断公羊生殖组织中的 B. ovis 时灵敏度略高。然而,不同核酸提取方案的灵敏度差异并不显著。大多数公羊只有一个生殖组织样本显示出阳性的 PCR 结果,因此建议从每只动物身上采集多个样本。要使分子检测诊断公羊附睾炎的灵敏度接近血清学检测的灵敏度,还需要进一步的研究。
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Reliability of Molecular Tests in Diagnosing Ovine Brucellosis Caused by Brucella Ovis
Brucella ovis infects sheep and causes a clinical or subclinical disease characterized by genital lesions and reduced fertility in rams, placentitis and abortions in ewes, and increased perinatal mortality in lambs. As part of this study, analyses were conducted on a sheep farm near Belgrade (Serbia). Of the serological tests, indirect ELISA was performed. A total of 94 blood sera were analyzed, 33 from rams and 61 from ewes. The results showed 23 (69.7%) positive findings in rams and 2 (3.3%) positive findings in ewes, with an overall prevalence of 25.4% bounded by a 95% confidence interval. Bruce-ladder multiplex PCR and Real time PCR were performed on 19 serologically positive rams and two serologically positive ewes. The results indicate a slightly higher sensitivity of Real time PCR compared to conventional PCR in diagnosing B. ovis from the reproductive tissues of rams. However, the differences in sensitivity between different nucleic acid extraction protocols were not significant. Most rams showed a positive PCR result in only one sample of reproductive tissue, suggesting the recommendation to take multiple samples from each animal. Further research is needed to bring the sensitivity of molecular tests in diagnosing ram epididymitis closer to the sensitivity of serological tests.
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