Ruiz-Cabello Jesús, Sevilla Iker A, Olaizola Ekine, Bezos Javier, Miguel-Coello Ana B, Muñoz-Mendoza Marta, Beraza Marta, Garrido Joseba M, Izquierdo-García Jose L
BRTA Basque Research and Technology Alliance, CIC biomaGUNE Center for Cooperative Research in Biomaterials, Donostia, Gipuzkoa, Spain.
Departamento de Química en Ciencias Farmacéuticas, Universidad Complutense de Madrid. Facultad de Farmacia, Madrid, Spain.
Transbound Emerg Dis. 2022 Jul;69(4):e859-e870. doi: 10.1111/tbed.14365. Epub 2021 Nov 19.
Even though enormous efforts and control strategies have been implemented, bovine tuberculosis (TB) remains a significant source of health and socioeconomic concern. The standard method used in TB eradication programs for in vivo detection is the tuberculin skin test. However, the specificity of the tuberculin skin test is affected by infection with non-tuberculous mycobacteria or by vaccination. Thus, some animals are not correctly diagnosed. This study aimed first to identify a plasma metabolic TB profile by high-field (HF) nuclear magnetic resonance (NMR) spectroscopy and second measure this characteristic TB metabolic profile using low-field benchtop (LF) NMR as an affordable molecular technology for TB diagnosis. Plasma samples from cattle diagnosed with TB (derivation set, n = 11), diagnosed with paratuberculosis (PTB, n = 10), PTB-vaccinated healthy control (n = 10) and healthy PTB-unvaccinated control (n = 10) were analyzed by NMR. Unsupervised Principal Component Analysis (PCA) was used to identify metabolic differences between groups. We identified 14 metabolites significantly different between TB and control animals. The second group of TB animals was used to validate the results (validation set, n = 14). Predictive models based on metabolic fingerprint acquired by both HF and LF NMR spectroscopy successfully identified TB versus control subjects (Area under the curve of Receiver Operating Characteristic over 0.92, in both models; Confidence Interval 0.77-1). In summary, plasma fingerprinting using HF and LF-NMR differentiated TB subjects from uninfected animals, and PTB and PTB-vaccinated subjects who may provide a TB-false positive, highlighting the use of LF-NMR-based metabolomics as a complementary or alternative diagnostic tool to the current diagnostic methods.
尽管已经实施了巨大的努力和控制策略，但牛结核病（TB）仍然是一个重大的健康和社会经济问题来源。结核病根除计划中用于体内检测的标准方法是结核菌素皮肤试验。然而，结核菌素皮肤试验的特异性会受到非结核分枝杆菌感染或疫苗接种的影响。因此，一些动物没有得到正确诊断。本研究的目的首先是通过高场（HF）核磁共振（NMR）光谱确定血浆代谢性结核病谱，其次使用低场台式（LF）NMR测量这种特征性的结核病代谢谱，作为一种经济实惠的结核病诊断分子技术。对诊断为结核病的牛（衍生组，n = 11）、诊断为副结核病（PTB，n = 10）、接种PTB疫苗的健康对照（n = 10）和未接种PTB疫苗的健康对照（n = 10）的血浆样本进行了NMR分析。使用无监督主成分分析（PCA）来识别组间的代谢差异。我们确定了结核病动物和对照动物之间有14种代谢物存在显著差异。第二组结核病动物用于验证结果（验证组，n = 14）。基于HF和LF NMR光谱获得的代谢指纹的预测模型成功地识别了结核病患者与对照受试者（两个模型中受试者工作特征曲线下面积均超过0.92；置信区间0.77 - 1）。总之，使用HF和LF - NMR进行血浆指纹分析可区分结核病患者与未感染动物，以及可能提供结核病假阳性的PTB和接种PTB疫苗的受试者，突出了基于LF - NMR的代谢组学作为当前诊断方法的补充或替代诊断工具的应用。
