Spectroscopic detection and identification of infected cells with herpes viruses

Abstract

Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and Fourier transform infrared (FTIR) microspectroscopy were previously applied for the identification of various biological samples. In the present study, normal cells in culture and cells infected with herpes simplex virus type 2 (HSV-2) or varicella-zoster virus (VZV) were analyzed by MALDI-TOF and FTIR microscopy. Specific spectral biomarkers for rapid and reliable monitoring and identification of infected cells and probably for the discrimination between these viruses were searched. The results show consistent spectral peaks in all examined normal uninfected human fibroblast cells both in MALDI-T0F and FTIR microscopy. In HSV-2- or VZV-infected cells, two unique peaks for each appeared at m/z 5397 and 5813 or at m/z 3501 and 4951, respectively, in MALDI-TOF spectra. In addition, several peaks that appeared in control uninfected cells at the region m/z 13,000-20,000 disappeared completely in all examined infected samples. When these infected cells were examined by FTIR microscopy, a band at 859 cm(-1) in control uninfected cells was significantly shifted to 854 cm(-1) in both HSV2- and VZV-infected cells. In addition, phosphate levels were considerably increased in all infected cells compared to normal uninfected cells. These parameters could be used as a basis for developing a spectral method for the detection and identification of cells infected with herpes viruses.

FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts

“…This technique has been previously explored for different measurement modes in many biological systems [2,3] to become a powerful tool for the cell components analysis, such as nucleic acids [4] proteins [5] and membranes [6]. This technique has also been useful for complex biological systems, such as tissues [7] and microorganisms [3,8,9] because of its ability to analyze both qualitative and quantitative, molecular and structural indications for samples components.…”

Identification of different respiratory viruses, after a cell culture step, by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS)

“…At present, mass spectrometry has had limited application in clinical virology6232425. In this light, the main purpose of this study was the development of a new tool based on MALDI-TOF MS as a model to be applied to the identification of cultivable respiratory viruses after a cell culture step.…”

“…However, serology and polymerase chain reaction (PCR) based assays are specific target directed and thus could miss non-selected viruses. On the other hand, although not target directed and then potentially able to detect all the cultivable viruses, cell culture is time consuming, expensive and needs accessory assays and trained personnel for virus identification, such as viral antigens detection by enzyme immunoassays, immunofluorescence5678 or electron microscopy.…”

“…Despite many literature data about MALDI-TOF MS application in routine clinical microbiology, and in experimental approaches related to bacteria, parasites, yeasts and moulds616171819202122, very few reports are available about its application in virus research and laboratory diagnosis of viral infections6232425. In most of the studies, specific viral genome sequences are previously amplified by PCR and then the amplicons are analysed by MALDI-TOF MS15.…”

Identification of Contaminated Cells with Viruses, Bacteria, or Fungi by Fourier Transform Infrared Microspectroscopy

“…Its applications cover different disciplines including material science, forensics, biochemistry, biomedical science, and geochemistry [3][4][5][6][7][8][9]. This technique has been also proved to be sensitive for the identification of cancer cells [1,[10][11][12][13], stem cells [14,15], virally infected cells [16][17][18][19], and microorganisms [20,21].…”