客户采用我司硅羧基磁珠偶联裂解型Siphovirus噬菌体LPST100，快速捕获沙门氏菌，建立了一种快速、准确的RT-PCR沙门氏菌检测方法。


Chenxi Huang, Binti Youssouf Mahboubat, Yifeng Ding, Qile Yang, Jia Wang, Min Zhou, Xiaohong Wang,
Development of a rapid Salmonella detection method via phage-conjugated magnetic bead separation coupled with real-time PCR quantification,
LWT,
Volume 142,
2021,
111075,
ISSN 0023-6438,
https://doi.org/10.1016/j.lwt.2021.111075.
(http://www.sciencedirect.com.group21-s.aronip.com/science/article/pii/S0023643821002280)

Abstract: In this study, we used the lytic Siphovirus phage LPST10 to develop a rapid and accurate detection method for Salmonella. Salmonella cells were separated from complex food matrices through magnetic separation, then subjected to SYBR green-based real-time quantitative PCR detection. Bacterial concentrations of 102–106 (Colony forming unit per milliliter, CFU/mL), with 200 of μg LPST10 phage-MBs, and 15 min reactions at 37 °C were found to be optimal to allow the phage-magnetic beads to capture the Salmonella cells. The phage-magnetic beads specifically recognized Salmonella cells of different serotypes. Under optimal conditions, DNA was extracted from the resulting bacteria phage-magnetic bead complexes through either alkaline lysis with sodium dodecyl sulfate (SDS) or boiling. The detection assay was subsequently assessed in food matrices (Tryptic soy broth medium, milk, and lettuce). The detection limit reached <30 CFU/mL in the food matrices. The entire assay, including bacterial capture, DNA extraction, and real-time polymerase chain reaction (RT-PCR) detection, could be completed within 1.5 h. This approach using a lytic Siphovirus phage has potential for rapid, specific, and accurate detection of Salmonella enterica subsp.enterica in different food matrices.
Keywords: Magnetic beads; Real-time PCR (Quantitative PCR); Salmonella biorecognition agent; Salmonella Typhimurium; Siphovirus


摘要：本研究利用裂解型Siphovirus噬菌体LPST10建立了一种快速、准确的沙门氏菌检测方法。用磁分离法从复杂的食品基质中分离沙门氏菌细胞，然后进行SYBR-green实时定量PCR检测。细菌浓度为102–106（每毫升菌落形成单位，CFU/mL），200μg LPST10噬菌体MBs，37℃反应15分钟°发现C是最适合让噬菌体磁珠捕捉沙门氏菌细胞的。噬菌体磁珠能特异识别不同血清型的沙门氏菌细胞。在最佳条件下，通过十二烷基硫酸钠（SDS）碱解或煮沸的方法从细菌噬菌体磁珠复合物中提取DNA。随后在食物基质（胰蛋白酶大豆肉汤培养基、牛奶和莴苣）中评估检测试验。食品基质的检出限<30cfu/mL。整个检测，包括细菌捕获、DNA提取和实时聚合酶链反应（RT-PCR）检测，可在1.5h内完成。这种利用裂解性Siphovirus噬菌体的方法有可能快速、特异、准确地检测不同食物基质中的肠道沙门氏菌亚种。

关键词：磁珠；实时定量PCR；沙门氏菌生物识别剂；鼠伤寒沙门菌；星状病毒