The multivariate detection limit for Mycoplasma pneumoniae as determined by nanorod array-surface enhanced Raman spectroscopy and comparison with limit of detection by qPCR.
The multivariate detection limit for Mycoplasma pneumoniae as determined by nanorod array-surface enhanced Raman spectroscopy and comparison with limit of detection by qPCR.
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DOI：
10.1039/c4an01141d
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发表时间：
2014-12-21
期刊：
影响因子：
--
通讯作者：
Krause DC
中科院分区：
文献类型：
--
作者：
Henderson KC;Sheppard ES;Rivera-Betancourt OE;Choi JY;Dluhy RA;Thurman KA;Winchell JM;Krause DC
肺炎支原体是人类呼吸道的一种无细胞壁细菌病原体，占社区获得性肺炎的20%。目前，用于检测和基因分型的标准是定量聚合酶链反应（qPCR），其可以表现出优异的灵敏度，但缺乏标准化，并且对于广泛的床旁使用具有有限的实用性。我们之前描述了一种能够检测M的Ag纳米棒阵列-表面增强拉曼光谱（NA-SERS）生物传感平台。在模拟和真实临床咽拭子样本中检测肺炎克雷伯氏菌的特异性和灵敏度具有统计学显著性。我们在这里报告，样品制备的差异影响支原体细胞的完整性NA-SERS分析，这反过来又影响所得的光谱。我们已经建立了一个多变量检测限（MDL）使用NA-SERS的M。肺炎完整细胞样品制备。使用国际纯粹与应用化学联合会（IUPAC）推荐的用于分析多变量数据集的方法，我们发现当以CFU/ml和DNA浓度（fg）表示时，qPCR的检测限比NA-SERS高约10倍。而完整M. pneumoniae为5.3 ± 1.0个基因组当量（cells/μl）。相比之下，一组平行样本的qPCR检测限为2.5 ± 0.25个细胞/μl。因此，对于某些标准度量，NA-SERS提供了M的多变量检测限。肺炎链球菌的抗体与通过qPCR测定的抗体基本相同。
Mycoplasma pneumoniae is a cell wall-less bacterial pathogen of the human respiratory tract that accounts for up to 20% of community-acquired pneumonia. At present, the standard for detection and genotyping is quantitative polymerase chain reaction (qPCR), which can exhibit excellent sensitivity but lacks standardization and has limited practicality for widespread, point-of-care use. We previously described a Ag nanorod array-surface enhanced Raman spectroscopy (NA-SERS) biosensing platform capable of detecting M. pneumoniae in simulated and true clinical throat swab samples with statistically significant specificity and sensitivity. We report here that differences in sample preparation influence the integrity of mycoplasma cells for NA-SERS analysis, which in turn impacts the resulting spectra. We have established a multivariate detection limit (MDL) using NA-SERS for M. pneumoniae intact-cell sample preparations. Using an adaptation of International Union of Pure and Applied Chemistry (IUPAC)-recommended methods for analyzing multivariate data sets, we found that qPCR had roughly 10× better detection limits than NA-SERS when expressed in CFU/ml and DNA concentration (fg). However, the NA-SERS MDL for intact M. pneumoniae was 5.3 ± 1.0 genome equivalents (cells/μl). By comparison, qPCR of a parallel set of samples yielded a limit of detection of 2.5 ± 0.25 cells/μl. Therefore, for certain standard metrics NA-SERS provides a multivariate detection limit for M. pneumoniae that is essentially identical to that determined via qPCR.
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