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对DNA度和细菌病原体PCR检测的比较,当使用两个DNA提取套件用于鼻/鼻样本时

Dam Khan¹, Shola-Able Thomas¹, Peggy-Estelle Tientcheu¹

¹Medical Research Council Unit The Gambia at the London School of Hygiene & Tropical Medicine, Banjul, The Gambia.

|August 3, 2023

概括

与DNeasy PowerLyzer PowerSoil Kit (Kit DP) 相比,DNeasy PowerLyzer PowerSoil Kit (Kit DPP) 的DNA度更高,病原体检测能力也相对较高. 这表明Kit DPP可能会改善鼻/鼻样本中少量细菌的恢复.

科学领域:

微生物学微生物学
分子生物学分子生物学
临床诊断临床诊断临床诊断

背景情况:

鼻/耳 (NP/OP) 无症状携带人类病原体需要准确的鉴定.
提取DNA对于培养独立的病原体检测方法至关重要.
DNA提取套件组件的变化可能会影响测试性能.

研究的目的:

为了比较两个DNA提取套件的DNA产量和病原体检测性能:DNeasy PowerSoil Kit (Kit DP) 和DNeasy PowerLyzer PowerSoil Kit (Kit DPP). 这两种DNA提取套件的DNA产量和病原体检测性能.
评估套件差异对NP/OP样本中Streptococcus pneumoniae,Haemophilus influenzae和Neisseria meningitidis的检测的影响.

主要方法:

从103个NP/OP样本中提取了DNA,使用了Kit DP和Kit DPP.
使用Qubit 2.0光计量DNA度.
实时聚合酶连锁反应 (RT-PCR) 用于检测S. pneumoniae,H. influenzae和N. meningitidis,使用Bland-Altman分析对Ct值达成一致.

主要成果:

与DPP套件 (884.9 ng/ml) 相比,DPP套件的平均DNA度 (1235.6 ng/ml) 显著更高 (p=0.002).
检测器件之间的病原体检测有很高的一致性 (>82%),Kit DP产生更高的Ct值 (表明DNA度或检测效率较低).
对于H. influenzae和N. meningitidis. 的90%以上的样本,获得了可复制的PCR结果.

结论:

来自Kit DPP的更高的DNA产量可能会提高NP/OP样本中低丰度细菌的检测.
两种套件都显示出很好的一致性,用于检测常见的NP/OP病原体,特别是阴性细菌.
在使用不同DNA提取方法的研究中比较结果时,应考虑套件之间的Ct值的变化.

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