PDF(Pubmed)
Abstract：
Rapid expansion of the probiotics industry demands fast, sensitive, comprehensive, and low-cost strategies for quality assessment. Here, we introduce a culture-free, one-cell-resolution, phenome-genome-combined strategy called Single-Cell Identification, Viability and Vitality tests, and Source-tracking (SCIVVS). For each cell directly extracted from the product, the fingerprint region of D2O-probed single-cell Raman spectrum (SCRS) enables species-level identification with 93% accuracy, based on a reference SCRS database from 21 statutory probiotic species, whereas the C-D band accurately quantifies viability, metabolic vitality plus their intercellular heterogeneity. For source-tracking, single-cell Raman-activated Cell Sorting and Sequencing can proceed, producing indexed, precisely one-cell-based genome assemblies that can reach ~99.40% genome-wide coverage. Finally, we validated an integrated SCIVVS workflow with automated SCRS acquisition where the whole process except sequencing takes just 5 h. As it is >20-fold faster, >10-time cheaper, vitality-revealing, heterogeneity-resolving, and automation-prone, SCIVVS is a new technological and data framework for quality assessment of live-cell products.
摘要：
益生菌行业的快速扩张需要快速，敏感,全面,和低成本的质量评估策略。这里,我们引入了无文化，单细胞分辨率，表型-基因组组合策略称为单细胞鉴定，活力和活力测试，和来源追踪(SCIVVS)。对于直接从产品中提取的每个细胞，D2O探测的单细胞拉曼光谱（SCRS）的指纹区域能够以93％的准确率进行物种级识别，基于来自21种法定益生菌物种的参考SCRS数据库，而C-D波段准确地量化了生存能力，代谢活力加上它们的细胞间异质性。对于源跟踪，单细胞拉曼激活的细胞分选和测序可以进行，生产索引，精确的基于单细胞的基因组组装，可以达到~99.40%的全基因组覆盖率。最后,wevalidedanintegratedSCIVVSworkflowwithautomaticallySCRSacquisitionwherethewholeprocessexceptsequencingtakesjust5h.Asitis>20-foldhigher,>10倍便宜，活力-揭示，异质性解决，易于自动化，SCIVVS是一种用于活细胞产品质量评估的新技术和数据框架。
