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客戶采用我司硅羧基磁珠偶聯(lián)裂解型Siphovirus噬菌體LPST100，快速捕獲沙門氏菌，建立了一種快速、準(zhǔn)確的RT-PCR沙門氏菌檢測(cè)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噬菌體LPST10建立了一種快速、準(zhǔn)確的沙門氏菌檢測(cè)方法。用磁分離法從復(fù)雜的食品基質(zhì)中分離沙門氏菌細(xì)胞，然后進(jìn)行SYBR-green實(shí)時(shí)定量PCR檢測(cè)。細(xì)菌濃度為102–106（每毫升菌落形成單位，CFU/mL），200μg LPST10噬菌體MBs，37℃反應(yīng)15分鐘°發(fā)現(xiàn)C是最適合讓噬菌體磁珠捕捉沙門氏菌細(xì)胞的。噬菌體磁珠能特異識(shí)別不同血清型的沙門氏菌細(xì)胞。在最佳條件下，通過十二烷基硫酸鈉（SDS）堿解或煮沸的方法從細(xì)菌噬菌體磁珠復(fù)合物中提取DNA。隨后在食物基質(zhì)（胰蛋白酶大豆肉湯培養(yǎng)基、牛奶和萵苣）中評(píng)估檢測(cè)試驗(yàn)。食品基質(zhì)的檢出限<30cfu/mL。整個(gè)檢測(cè)，包括細(xì)菌捕獲、DNA提取和實(shí)時(shí)聚合酶鏈反應(yīng)（RT-PCR）檢測(cè)，可在1.5h內(nèi)完成。這種利用裂解性Siphovirus噬菌體的方法有可能快速、特異、準(zhǔn)確地檢測(cè)不同食物基質(zhì)中的腸道沙門氏菌亞種。

關(guān)鍵詞：磁珠；實(shí)時(shí)定量PCR；沙門氏菌生物識(shí)別劑；鼠傷寒沙門菌；星狀病毒