Virus infection of a human cell was determined only 3 h after invagination. We used viral vector Ad-CMV-control (AdC), which lacks the E1 gene coding for early polypeptide 1 (E1). AdC can replicate in human embryonic kidney 293 (HEK293) cells into which the E1 gene has been transfected. According to partial least-square regression discriminant analysis, it was assumed that two kinds of reaction take place in the cell during viral invasion. The first response of the cell was determined 3 h after the virus invasion, and the second one was determined ∼9 h later. The first one seems to be due to compositional changes in DNA. Analysis of large-scale datasets strongly indicated that the second reaction can be attributed to a reduction in protein concentration or uptake of phenylalanine into the nucleus.
The present study demonstrates that Raman spectroscopy is a powerful tool for the detection of virus-infected cells. Adenovirus infection of human embryonic kidney 293 cells was successfully detected at 12, 24, and 48 h after initiating the infection. The score plot of principal component analysis discriminated the spectra of the infected cells from those of the control cells. The viral infection was confirmed by the conventional immunostaining method performed 24 h after the infection. The newly developed method provides a fast and label-free means for the detection of virus-infected cells.
Raman spectroscopy is a promising tool for detection of virus infection in live cells. In the present study, we
demonstrate its feasibility to observe dynamic reaction of the live cell infected by virus. The Raman spectra of the
adenovirus infected live cell (293 HEK) are analyzed by comparing with those of control cells. Principal
component analysis (PCA) is employed also to analyze the spectra in detail. A band at 1650 cm-1 increases its
intensity in the spectra measured at 24 hours after the virus infection. The infection of the virus is also examined
by immune-staining and transmission electron microscope (TEM), and the virus infection is confirmed with these
method also. It should be noted that the present technique does not require specifying the type of virus in
advance.
Problem of viruses is very actual for nowadays. Some viruses, which are responsible for human of all tumors, are about
15 %. Main purposes this study, early detection virus in live cell without labeling and in the real time by Raman
spectroscopy. Micro Raman spectroscopy (mRs) is a technique that uses a Raman spectrometer to measure the spectra of microscopic samples. According to the Raman spectroscopy, it becomes possible to study the metabolites of a live
cultured cell without labeling. We used mRs to detect the virus via HEK 293 cell line-infected adenovirus. We obtained
raman specters of lives cells with viruses in 24 hours and 7 days after the infection. As the result, there is some
biochemical changing after the treatment of cell with virus. One of biochemical alteration is at 1081 cm-1. For the clarification result, we use confocal fluorescent microscopy and transmission electron microscopy (TEM).
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