• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Tropical Animal Health and Production >Improvement of sensitivity for Mycobacterium avium subsp. paratuberculosis (MAP) detection in bovine fecal samples by specific duplex F57/IC real-time and conventional IS900 PCRs after solid culture enrichment
【24h】

Improvement of sensitivity for Mycobacterium avium subsp. paratuberculosis (MAP) detection in bovine fecal samples by specific duplex F57/IC real-time and conventional IS900 PCRs after solid culture enrichment

机译：提高对鸟分枝杆菌亚种的敏感性。固体培养物富集后，通过特定的双工F57 / IC实时和常规IS900 PCR在牛粪便样品中检测肺结核旁（MAP）

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of Johne's disease in ruminants and a probable pathogen of Crohn's disease in humans. Accurate, cost-effective, and time-relevant diagnostics are the basis for efficient control programs. This study was conducted as an attempt to re-evaluate MAP detection improvement by coupling solid media enrichment to a more specific IS900 conventional PCR and a very specific F57/IC real-time PCR. In a spiking experiment, we investigated the improvement of molecular-based MAP detection in feces after a culture-based enrichment step into Herrold's egg yolk media with mycobactin J (HEYM-MJ) for different time intervals, when compared to traditional culture. Detection limit of culture was 0.33 x 10(4) bacteria x g(-1) (33 CFU g(-1)), while that of IS900 PCR when coupled with an enrichment step for 2, 4, and 6 weeks was 0.33 x 10(5) (0.33 x 10(3) CFU g(-1)), 0.33 x 10(4) (33 CFU g(-1)), and 33 (> 3.3 CFU g(-1)) bacteria x g(-1), respectively. Whereas the detection limits of F57/IC real-time PCR after the enrichment step for the same time intervals were 0.33 x 10(5) (0.33 x 10(3) CFU g(-1)), 0.33 x 10(3) (3.3 CFU g(-1)), and 33 (> 3.3 CFU g(-1)) bacteria x g(-1), respectively. Altogether, enrichment of bovine fecal samples into solid media increased the sensitivity of specific molecular detection of MAP using IS900 conventional PCR and duplex F57/IC real-time PCR and offers an expedited and accurate alternative for MAP detection in bovine feces. Validation of these results is further recommended using field bovine fecal samples.
机译：鸟分枝杆菌亚种副结核病（MAP）是反刍动物中约翰氏病的病原体，也是人类克罗恩氏病的可能病原体。准确，具有成本效益和与时间相关的诊断是有效控制程序的基础。进行这项研究是为了尝试通过将固体培养基富集耦合到更特定的IS900常规PCR和非常特定的F57 / IC实时PCR来重新评估MAP检测的改进。在一个尖峰实验中，与传统培养相比，我们研究了在不同时间间隔内，使用霉菌素J（HEYM-MJ）在基于Herrold的蛋黄培养基中进行基于文化的富集步骤后，粪便中基于分子MAP检测的改进。培养物的检出限为0.33 x 10（4）细菌xg（-1）（33 CFU g（-1）），而与900、2、4和6周的富集步骤偶联的IS900 PCR的检出限为0.33 x 10 （5）（0.33 x 10（3）CFU g（-1）），0.33 x 10（4）（33 CFU g（-1））和33（> 3.3 CFU g（-1））细菌xg（- 1）。富集步骤之后相同时间间隔的F57 / IC实时PCR的检出限为0.33 x 10（5）（0.33 x 10（3）CFU g（-1）），0.33 x 10（3）（ 3.3个CFU g（-1））和33个（> 3.3 CFU g（-1））细菌xg（-1）。总而言之，将牛粪便样品富集到固体培养基中可以提高使用IS900常规PCR和双工F57 / IC实时PCR进行MAP的特异性分子检测的灵敏度，并为牛粪中MAP的检测提供了一种快速而准确的替代方法。进一步建议使用野牛粪便样品验证这些结果。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号