比较三种不同 PCR 方案检测黄病毒属的效果。
Comparison of three different PCR protocols for the detection of ferlaviruses.
机构信息
Laboklin GmbH & Co. KG, Steubenstraße 4, 97688, Bad Kissingen, Germany.
School of Veterinary Medicine, Murdoch University, Murdoch, Western Australia, 6150, Australia.
出版信息
BMC Vet Res. 2019 Aug 6;15(1):281. doi: 10.1186/s12917-019-2028-0.
BACKGROUND
Ferlaviruses are important pathogens in snakes often associated with respiratory and neurological disease. The detection of ferlaviral RNA by PCR is considered to be the most reliable method for the diagnosis of infection. The PCRs that have been used most commonly for this purpose have not been properly assessed to determine their sensitivity, specificity and ability to detect the known genetic diversity of this group of viruses. The aim of this study was to compare three published PCR protocols so that a single method could be recommended to laboratories that perform this testing.
RESULTS
Comparisons were carried out using cell culture isolates and tissues from snakes infected with specific virus genotypes. A single round PCR targeting a short segment of the viral polymerase (L) gene provided the highest sensitivity and specificity, and detected isolated ferlaviruses from all four described genogroups, as well as from tissues of infected snakes.
CONCLUSION
A broadly-reactive PCR for the detection of all known ferlaviruses was found to provide a good combination of detection limit, specificity and speed. Based on these criteria, this method is recommended for the diagnosis of ferlavirus infections.
背景
铁蛋白病毒是蛇类中的重要病原体，常与呼吸道和神经系统疾病有关。通过 PCR 检测铁蛋白病毒 RNA 被认为是诊断感染的最可靠方法。为此目的而使用的最常见的 PCR 尚未经过适当评估，以确定其灵敏度、特异性和检测该病毒组已知遗传多样性的能力。本研究的目的是比较三种已发表的 PCR 方案，以便向进行此项检测的实验室推荐一种单一方法。
结果
使用针对特定病毒基因型感染的蛇的细胞培养物分离物和组织进行了比较。针对病毒聚合酶（L）基因短片段的单轮 PCR 提供了最高的灵敏度和特异性，并检测到来自所有四个描述的基因群的分离铁蛋白病毒，以及来自感染蛇的组织。
结论
发现一种针对所有已知铁蛋白病毒的广谱反应性 PCR 具有良好的检测限、特异性和速度组合。基于这些标准，建议将该方法用于铁蛋白病毒感染的诊断。
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