开发和评估滴滴数字PCR试验,以检测人类全血中的布鲁塞拉菌
Jiayin Liu1,2, Zhichun Song2, Na Ta3
1National Key Laboratory of Intelligent Tracing and Forecasting for Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
PLoS neglected tropical diseases
|June 2, 2023
概括
滴滴数字PCR (ddPCR) 提供了一种灵敏而准确的方法来量化全血中的布鲁塞拉DNA,有助于诊断和预测血病. 这项技术在检测病原体方面,与定量PCR (qPCR) 相比,表现优越.
科学领域:
- 分子生物学分子生物学
- 传染性疾病 传染性疾病
- 生物技术是生物技术.
背景情况:
- 动物传播疾病 - - 动物病菌 - - 由于其在养动物中日益流行,对公众健康构成重大挑战.
- 在流行地区,准确及及时诊断人类血病仍然很困难,这会影响患者的管理和结果.
研究的目的:
- 建立和评估滴滴数字PCR (ddPCR) 方法,以精确量化人类全血样本中的布鲁塞拉DNA.
- 评估开发的ddPCR测定对人类血病的诊断,预后和治疗价值.
主要方法:
- 为针对布鲁塞拉bcsp31基因的ddPCR试验优化原料和探针度和回火温度.
- 测定灵敏度,特异性和可重复性的评估,使用来自确诊和疑似牛病例的全血样本.
- 对ddPCR与定量PCR (qPCR) 进行比较分析,以检测布鲁塞拉DNA并监测随访患者的治疗反应.
主要成果:
- 优化的ddPCR试验显示检测极限为1.87副本/反应,具有很高的重复性.
- 在血清凝结试验阳性患者中,ddPCR实现了比qPCR (75.4%) 更高的阳性检测率 (88.5%).
- ddPCR在57.6%的疑似血清阴性样本中发现了布鲁塞拉DNA,明显高于qPCR的36.3%.
结论:
- 已建立的ddPCR测定方法对于在全血样本中量化布鲁塞拉DNA是准确的.
- ddPCR显示出潜在的作为一个有价值的工具,用于诊断和预后的人类牛病.
- 该测试能够追踪DNA负载的变化,可以为治疗管理和疾病监测提供信息.
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