蛋白酶K预处理的基于CONA的ELISA测定:一种新的尿液LAM检测策略用于结核病诊断
Huan Huang1,2, Rong Qu2,3, Kang Wu2
1Shanghai Key Laboratory of Atmospheric Particle Pollution Prevention (LAP3), Department of Environmental Science and Engineering, Fudan University, Shanghai, China.
Frontiers in microbiology
|September 11, 2023
概括
使用康卡纳瓦林A (ConA) 和蛋白酶K的新测定方法有效地检测尿液中的利波阿拉比诺南 (LAM),改善结核病 (TB) 诊断. 这种方法对敏感和特定的结核病检测有希望,解决了当前测试的局限性.
科学领域:
- 免疫诊断 免疫诊断 免疫诊断 免疫诊断
- 菌根细菌的研究研究.
- 生物化学 生化学
背景情况:
- 甲 (Lipoarabinomannan,LAM) 是Mycobacterium tuberculosis (Mtb) 的关键细胞壁糖脂,也是结核病 (TB) 诊断的潜在生物标志物.
- 现有的商业尿液LAM测试缺乏足够的灵敏度,因此需要改进诊断策略.
- 开发新的LAM检测方法对于弥合目前结核病诊断差距至关重要.
研究的目的:
- 开发一种新型的诊断试验,用于检测尿样中的利波阿巴比诺南 (LAM).
- 评估使用各种菌根菌株和临床样本开发的试验的诊断性能.
- 评估蛋白酶K预处理对测定对结核病检测的敏感性和特异性的影响.
主要方法:
- 基于康卡纳瓦林A (ConA) 的ELISA试验被开发并使用蛋白酶K预处理进行优化.
- 对ManLAM的测试检测极限 (LoD) 确定为6 ng/ml.
- 用细菌菌株和71个来自结核病患者,健康对照者和密切接触者的临床尿样来评估诊断性能.
主要成果:
- 基于ConA的ELISA显示出对Mtb和M. bovisBCG的强烈反应性,对M. smegmatis的中间反应性,以及对非真菌细菌的弱反应性.
- 蛋白酶K的预处理显著增强了对LAM的抗体反应性.
- 该试验将结核病患者与对照患者区分开来,敏感度为43.8% (TB+HIV+) 和37.5% (TB+HIV-),特异性为100.0%,AUC为0.74和0.82.
结论:
- 康卡纳瓦林A可以有效地与抗体配对,用于检测Lipoarabinomannan (LAM).
- 蛋白酶K预处理是通过恢复抗体活性来提高LAM检测的灵敏度的一种有价值的方法.
- 这种新型测试显示出作为结核病 (TB) 的改进诊断工具的潜力.
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