An Imaging Technique For Detection Of Virus Assisted With Nanofluorescent Materials

Background:Viruses are the most significant pathogens responsible for the high morbidity and mortality of diseases worldwide,especially since the 2019 pandemic of the new coronavirus(SARS-Co V-2),which has had a huge impact on human health and society。So far,there are no very effective drugs or methods to treat and prevent the spread of disease.Therefore,the establishment of rapid,reliable and relatively low-cost virus detection technologies is an important means of controlling the spread of the virus.Currently existing virus detection technologies focus on the detection of nucleic acids and antigens of viruses or antibodies produced as a result of viruses,however these methods also have limitations,that’s why so much focus on new type of virus detection technologies.which developed based on the spectral characteristics of viruses,such as surface-enhanced Raman spectroscopy(SERS),have been found to be very promising methods for virus detection and some of these have been successfully developed and applied,such as SERS techniques have been used for the direct detection of dengue virus,human immunodeficiency virus,H1N1 influenza virus,avian influenza virus and others.These novel virus detection methods,which differ from conventional techniques,are important and have applications for the prevention and monitoring of virus epidemics.In this study,we developed and established a novel virus detection technique combining rare earth nanofluorescent materials with an ultra-weak biophotonic imaging system,which may be of potential value in virus screening and surveillance.Methodology:By mixing two nanomaterials and a glycerol buffer,a stable colloidal weak light source was developed that fluoresces under 230 nm UV excitation and can assist in the dynamic imaging of viral samples.The key methodology and evaluation process for this developed technology includes:(1)the design and development of a quartz glass nine-well plate as a sample test plate,where the cylindrical sample well volume is approximately 15μl with a smooth inner surface and good light reflectivity;(2)the technical evaluation using three different concentrations of lentivirus(RNA virus),adenovirus(DNA virus)and adeno-associated virus(DNA virus);(3)the evaluation of the sample by means of an ultra-weak bio-light imaging system to detect dynamic changes in the intensity of the weak light presented by the test sample;(4)the sensitivity and specificity of the technique was assessed using subject operating characteristic curve(ROC)analysis.Results:Two types of nanomaterials that fluoresce yellow and green under UV excitation were selected and configured into a colloidal solution using 60%glycerol buffer and finally filtered appropriately.The results showed that the colloidal solution had a homogeneous particle fraction and showed a stable and characteristic weak luminescence decay process upon excitation by UV light.Compared to the control,the lentivirus(RNA virus)samples at different concentrations(1.87×10~5,1.87×10~4and 1.87×10~3TU/m L,respectively)showed a significant concentration-dependent decrease compared to the control,but such a decrease was only found in the assessment of adeno-associated viruses(DNA viruses)at high titres(4.7×10~6VG/m L),while low and medium concentrations(4.7×10~5VG/m L,respectively)were found.(4.7×10~5and 4.7×10~4VG/m L,respectively)showed no significant performance.Furthermore,no significant decreases were found for adeno-associated viruses(DNA viruses)at different concentrations(1.4×10~6,1.4×10~5and 1.4×10~4PFU/m L,respectively).In addition,a certain concentration of the non-luminous bacterium Staphylococcus aureus enhanced the weak light intensity of the nanoluminescent solution.Taking control as a reference,we evaluated the sensitivity and specificity of high concentration of virus samples,and found that the area under the curve(AUC)was 0.7560,while the sensitivity and specificity were 0.6154 and0.7714,respectively.Conclusion:These results suggest that viruses absorb a specific spectrum of faint luminescence under rare earth nanocolloid solution conditions,which is related to the concentration and type of virus,and the mechanism of which deserves further exploration.The preliminary results of this study provide an important basis for the further development of this technology and new ideas for the extended development of related technologies.