Foto del docente

Mara Mirasoli

Associate Professor

Department of Chemistry "Giacomo Ciamician"

Academic discipline: CHIM/01 Analytical Chemistry

Research

Keywords: bio- chemiluminescence biosensors immunoassay imaging electronic nose nucleic acids hybridization methods point-of-care testing

- Development of chemiluminescent enzyme assays, immunoassays and gene probe assays, for the quantitative analysis of molecules of interest in the medical, alimentary and environmental fields.

- Development of new portable devices for multiplexed detection based on chemiluminescence and using microfabrication technology and microfluidics

- Development of ultrasensitive immunohistochemistry and in situ hybridization techniques with luminescent detection for the simultaneous quantitative localization of proteins and/or nucleic acid sequences in cells or tissue sections

- Application of detection techniques based on chemiluminescence measurements to field-flow fractionation (FFF) and development of hybrid FFF systems employing antigen-antibody interactions.

- Development of methods based on the electronic nose technology for the characterization of natural products and cosmetics, based on their olfactive fingerprint.

 

- Development of chemiluminescent immunoassays and gene probe assays, for the quantitative analysis of molecules of interest in the medical, alimentary and environmental fields. Recently, chemiluminescence enzyme immunoassays were developed for the quantification of: (1) tauroursodeoxycholic acid in oral fluid for diagnostic purposes; (2) estradiol, estrone and estriol in wastewater samples; (3) the protein Cry1Ab in maize samples in order to detect the presence of geneticallay modified Bt maize; (4) aflatoxin M1 in milk samples; (5) zeranol, trenbolone and nortestosterone in bovine urine. PCR-ELISA assays in innovative microtiter plate formats were developed for detection and genotyping of Papillomavirus in biological samples.

- Development of new portable devices for multiplexed detection based on chemiluminescence and using microfabrication technology and microfluidics. A simple and versatile analytical device has been realized employing a cooled ultrasensitive portable CCD camera in a contact imaging configuration. The device is suitable for point-of-care applications and it is designed to perform, even simultaneously, different types of bioassays. A miniaturized multiplex biosensor exploiting a microfluidic oligonucleotide array and chemiluminescence (CL) lensless imaging detection has been developed for parvovirus B19 genotyping. Lateral flow immunoassays (LFIA) with chemiluminescence were developed for detecting: (1) fumonisin B1 in maize flour samples; (2) explosive traces in environmental samples.

- Development of new silica nanoparticles loaded with thermochemiluminescent compounds. These nanoparticles appear to be very promising for their implementation in lab-on-chip devices, since the trigger of the chemical luminescent signal does not require the addition of reagents and it is triggered simply by the application of a thermal shock through integrated microheaters.

- Development of ultrasensitive immunohistochemistry and in situ hybridization techniques with luminescent detection for the simultaneous quantitative localization of proteins and/or nucleic acid sequences in cells or tissue sections. Methods combining luminescent IHC and ISH were developed for the co-localization of the melanocitic marker HMB-45 and human papillomavirus (HPV) DNA in melanoma tissue sections and for the co-localization of the protein p16INK4a (also its quantification) and HPV DNA in uterine cervix lesions; a chemiluminescent method for the localization and quantification of Parvovirus B19 nucleic acids in infected cells employing peptide nucleic acid (PNA) probes was developed. Recently, quantitative methodologies exploiting ultrasensitive chemiluminescence detection of 5-bromo-2′-deoxyuridine (BrdU) have been developed: a CL microscope imaging assay to evaluate BrdU labelling at single-cell level and a CL dot-blot assay to measure the amounts of DNA produced in the course of an in vitro infection of proliferating cells.

- Application of detection techniques based on chemiluminescence measurements to field-flow fractionation (FFF) and development of hybrid FFF systems employing antigen-antibody interactions. A method for automated preparation of plasma from whole blood and on-line quantification of alkaline phosphatase activity in plasma by chemiluminescence detection has been developed to proof the possibility to exploit FFF for biological sample pre-analytical treatment in point-of-care testing devices. FFF has been coupled to electronic nose detection to allow pathogen bacteria detection in milk samples and differentiation between live and dead bacterial cells. A method for increasing the selectivity of gravitational FFF in cell sorting has been developed by modifying the channel accumulation wall with antibodies, able to selectively slow down the elution of specific cells.

- Development of methods based on the electronic nose technology for the characterization of natural products and cosmetics, based on their olfactive fingerprint. We developed e-nose-based methods for: (1) the characterization of green tea based on regional origin and state of conservation; (2) characterization of aromatherapy creams based on the type and amount of essential oil and on the state of conservation.

- Isotope ratio mass spectrometry (13C/12C IRMS) for the development of new “breath tests” for rapid non invasive diagnosis of liver and gastrointestinal diseases. Breath tests were employed for monitoring the motilities and functionalities of the gastrointestinal system in human space mission simulations Mars500.

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