Quantitative detection of the Ralstonia solanacearum species complex in soil by qPCR combined with a recombinant internal control strain.
Wei ChenJun-Wei ZhangBi-Xia QinHui-Ting XieZhi ZhangXiu-Ze QiaoShan-Kui LiMuhammad AsifSong GuoLi-Xian CuiPei-Pei WangLi-Hong DongQing-Gang GuoWen-Jun JiangPing MaZhen-Yuan XiaCan-Hua LuLi-Qun ZhangPublished in: Microbiology spectrum (2023)
species complex (RSSC), plays a vital role in risk assessment, but meanwhile, precise quantification is difficult due to the poor purity and yield of the soil DNA retrieved. The internal sample process control (ISPC) strain RsPC we developed solved this problem and significantly improved the accuracy of quantification of RSSC in different soils. ISPC-based quantitative PCR detection is a method especially suitable for the quantitative detection of microbes in complex matrices (such as soil and sludge) containing various PCR inhibitors and for those not easy to lyse (like Gram-positive bacteria, fungi, and thick-wall cells like resting spores). In addition, the use of ISPC strains removes additional workload on the preparation of high-quality template DNA and facilitates the development of high-throughput quantitative detection techniques for soil microbes.
Keyphrases
- real time pcr
- loop mediated isothermal amplification
- risk assessment
- high throughput
- high resolution
- label free
- heavy metals
- escherichia coli
- cell free
- induced apoptosis
- microbial community
- wastewater treatment
- heart rate
- blood pressure
- human health
- multidrug resistant
- climate change
- cell death
- signaling pathway
- single cell
- mass spectrometry
- endoplasmic reticulum stress
- sensitive detection
- tandem mass spectrometry