通过减少无细胞DNA对Mycobacterium tuberculosis量化和唾液微生物群的混影响来减去背景
Charissa C Naidoo1,2, Rouxjeane Venter1, Francesc Codony3
1DSI-NRF Centre of Excellence for Biomedical Tuberculosis Research, South African Medical Research Council Centre for Tuberculosis Research, Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences, Stellenbosch University, PO Box 241, Cape Town, 8000, South Africa.
Scientific reports
|September 27, 2024
概括
核酸染料,如氧化 (PMA) 和PEMAX,有效地减少结核病诊断中的细胞外DNA. 这些染料改善了Mycobacterium检测和微生物组分析,特别是在化疗期间.
科学领域:
- 微生物学 微生物学
- 分子诊断学 分子诊断
- 基因组学就是基因组学.
背景情况:
- 细胞外DNA混结核病 (TB) 诊断和微生物组研究,特别是在接受化疗的患者中.
- 精确的DNA表征对于结核病诊断和了解相关的微生物群至关重要.
研究的目的:
- 评估核酸染料 (propidium monoazide [PMA],PEMAX) 和DNaseI在减少细胞外DNA中的有效性.
- 评估这些治疗方法对结核病患者的Mycobacterium tuberculosis complex (Mtb) DNA检测和微生物组概况的影响.
主要方法:
- 量化PCR (qPCR) 和针对16S rRNA基因的测序在Mtb稀释系列和人类唾液样本上进行.
- 在分子分析之前,样品被PMA,PEMAX或DNaseI处理.
- 从结核病患者 (A组和B组) 的唾液在化疗前后被分析.
主要成果:
- PMA和PEMAX显著降低了PCR检测到的Mtb DNA,表明在治疗前存在不完整的Mtb.
- 在58%的结核病阳性样本中,PEMAX通过测序增强了Mycobacterium检测.
- 染料改变了唾液微生物组中的α和β多样性指标,PEMAX揭示了与化疗相关的变化.
结论:
- PMA和PEMAX有效地减少了细胞外DNA,改善了结核病诊断中的病原体检测和微生物组分析.
- 佩马克斯对于检测化疗诱导的微生物社区转移特别有价值.
- 核酸染料是精确评估结核病患者的微生物群的重要工具,特别是在治疗期间.
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