Estimation of Mycobacterium avium subsp paratuberculosis Growth Parameters: Strain Characterization and Comparison of Methods
Estimation of Mycobacterium avium subsp paratuberculosis Growth Parameters: Strain Characterization and Comparison of Methods
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DOI：
10.1128/aem.05818-11
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发表时间：
2011-12-01
影响因子：
4.4
通讯作者：
Juste, Ramon A.
中科院分区：
文献类型：
--
作者：
Elguezabal, Natalia;Bastida, Felix;Juste, Ramon A.
鸟分枝杆菌亚种的生长速度。在补充有OADC（油酸、白蛋白、右旋糖、过氧化氢酶）、吐温80和分枝杆菌素J的7 H9培养基中通过不同方法评估副结核杆菌。通过湿重、浊度测量、活菌计数和定量PCR（ParaTB-Kuanti; F57基因）确定8 M的代次和最大比生长速率。鸟亚种副结核杆菌菌株（K10、2 E、316 F、81、445、764、22 G和OVICAP 49）。在生长曲线和计算参数中观察到应变间差异。定量方法在特定时间点对每种菌株给出了不同的结果。世代时间范围从活菌计数和qPCR的平均1.4天到湿重和浊度测定的约10天。湿重法、浊度法和ParaTB-Kuanti qPCR法彼此相关性最好。通常，通过活菌计数作为参考方法评估活力;鸟亚种副结核菌丛生问题和不可培养的M.鸟亚种我们得出结论，单拷贝基因的qPCR可以可靠地用于快速估计M。鸟亚种样本中的副结核菌数量。
The growth rate of Mycobacterium avium subsp. paratuberculosis was assessed by different methods in 7H9 medium supplemented with OADC (oleic acid, albumin, dextrose, catalase), Tween 80, and mycobactin J. Generation times and maximum specific growth rates were determined by wet weight, turbidometric measurement, viable count, and quantitative PCR (ParaTB-Kuanti; F57 gene) for 8 M. avium subsp. paratuberculosis strains (K10, 2E, 316F, 81, 445, 764, 22G, and OVICAP 49). Strain-to-strain differences were observed in growth curves and calculated parameters. The quantification methods gave different results for each strain at specific time points. Generation times ranged from an average of 1.4 days for viable count and qPCR to approximately 10 days for wet weight and turbidometry. The wet-weight, turbidometry, and ParaTB-Kuanti qPCR methods correlated best with each other. Generally, viability has been assessed by viable count as a reference method; however, due to M. avium subsp. paratuberculosis clumping problems and the presence of noncultivable M. avium subsp. paratuberculosis cells, we conclude that qPCR of a single-copy gene may be used reliably for rapid estimation of M. avium subsp. paratuberculosis bacterial numbers in a sample.