诊断方面的进展:分离Mycobacterium avium ssp. 这种细菌. 结核病并揭示了其与嵌套PCR的分子身份
Faranak Nouri1, Alireza Shahrjerdi2, Hossein Ali Zarnegarpour3
1Department of Microbiology, Faculty of Science, Islamic Azad University Arak Branch, Arak, Iran. 351157@gmail.com.
Cellular and molecular biology (Noisy-le-Grand, France)
|March 2, 2024
概括
这项研究从伊朗的ELISA阳性牛中分离了Mycobacterium avium亚种结核病 (MAP). 分子方法证实了MAP的存在和基因表达,有助于识别受感染的动物并减少经济损失.
科学领域:
- 兽医微生物学 兽医微生物学
- 牛的传染病 牛的传染病
- 分子诊断学 分子诊断学
背景情况:
- 结核病是由Mycobacterium avium亚种结核病 (MAP) 引起的,在伊朗的畜牧业中带来了重大的经济挑战.
- 准确识别和隔离MAP对于控制其传播和减轻经济损失至关重要.
- 目前的诊断方法需要验证,以便在MAP感染的群体中有效地实地应用.
研究的目的:
- 从阳性牛牛的牛奶和便样本中分离MAP.
- 为了确定孤立的MAP菌株的分子特征.
- 评估感染动物中关键的MAP相关基因的表达水平.
主要方法:
- 与酶相关的免疫吸收试验 (ELISA) 选了3700个牛血液样本.
- 在选择性介质上培养来自ELISA阳性牛的牛奶和便样本的细菌学培养.
- 酸固性染色 (Ziehl-Neelsen),16S rRNA PCR,IS900 PCR和嵌套PCR用于分子识别.
- 定量PCR (qPCR) 用于评估MAP特定基因的表达 (IS900,ISMAP02,F57,MAP2191,MAP4027) 的情况.
主要成果:
- 从90个ELISA阳性样本中成功培养出9个MAP分离物,所有样本都表现出酸性.
- 16S rRNA PCR证实了Mycobacterium的存在,而IS900 PCR证实了MAP的识别.
- 嵌套PCR证实了所有阳性样本,qPCR揭示了MAP相关基因在ELISA阳性样本中的显著更高的表达相比负面样本.
结论:
- 艾丽莎是一种有效的查工具,用于识别潜在的MAP感染牛.
- 细菌学培养与分子技术 (嵌套PCR) 结合,可提供MAP的可靠确认.
- 在ELISA阳性动物中MAP特异基因的高表达支持分子方法的诊断实用性.
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