Methods and systems for detection of biohazard signatures in complex clinical and environmental samples

A method for identifying presence of a selected biohazard signature in a sample, comprising:

• (a) exposing a sample to excitation radiation and reading resulting emission radiation for a plurality of wavelength bands from each of a plurality of locations on the sample;
• (b) on a location-by-location basis, performing a first level analysis to determine which emission radiation readings meet signal threshold requirements relative to background noise readings;
• (c) for each location having emission radiation readings meeting signal threshold requirements, performing a second level membership analysis to assign a class membership to individual location readings indicative of potential relevance to biohazard signature presence, wherein positive membership is assigned to locations when the readings for such locations are deemed closer to readings associated with samples known to contain the biohazard signature than to readings associated with samples known not to contain the biohazard signature;
• and
• (d) defining at least one minimum threshold spatial grouping criterion for locations with positive membership that is necessary to conclude that the sample is positive for presence of the biohazard signature;
• (e) performing at least one additional level of analysis comprising a determination of whether the sample includes positive membership locations that, taken together in conjunction with their relative spatial positions, meet the at least one minimum threshold spatial grouping criterion and if so, determining that the sample is positive for the presence of the biohazard signature,
• wherein the excitation radiation comprises radiation with wavelength selected from the group consisting of:
• (1) below 300 nanometers, (2) below 275 nm, and (3) below 250 nm,
• wherein the emission radiation readings comprise readings of native fluorescence emission radiation, and
• wherein the at least one minimum threshold spatial grouping criterion comprises use of positive membership for multiple locations and the spatial relationship of those locations.

A method for identifying presence of a selected biohazard signature in a complex sample, comprising:

• (a) providing excitation radiation onto a portion of a sample, wherein the excitation radiation is provided as a beam from a source within a housing, wherein the beam has a dimension that is smaller than a sample dimension;
• (b) receiving emission radiation, from the portion of the sample that received excitation radiation, onto at least one optical element which directs the emission radiation along at least one detection path within the housing;
• (c) directing excitation radiation to be incident upon different portions of a sample by relatively moving the beam and the sample;
• (d) for each of a plurality of different portions of the sample, detecting a group of emission signals, with each group comprising signals from a plurality of different wavelength bands, using at least one detector located within the housing at a location along the at least one detection path;
• (e) for the plurality of groups of emission signals associated with the plurality of different sample portions, distinguishing useful signal groups from unusable signal groups, where useful signal groups are those containing at least one signal having a strength that is greater than a strength of a corresponding background signal by a predefined amount, and wherein each useful signal group is associated with one of the plurality of difference sample portions;
• (f) providing both predetermined biohazard indicative signal information and predetermined non-biohazard indicative signal information related to the selected biohazard signature;
• (g) producing a biohazard indicative status for each useful signal group based at least in part upon a comparison of emission signal data for that group with the predetermined biohazard indicative signal information and predetermined non-biohazard indicative signal information;
• and
• (h) forming a biohazard indicative conclusion based, at least in part, on a combination of (1) biohazard indicative signal information for a plurality of useful signal groups, and (2) relative spacings between the portions of the sample associated with the plurality of useful signal groups having biohazard indicative signal information.

A method of identifying presence of a selected biohazard signature in a complex sample, comprising:

• (a) providing excitation radiation onto a portion of a sample, wherein the excitation radiation is provided as a beam from a source within a housing, wherein the beam has a dimension that is smaller than a sample dimension;
• (b) receiving emission radiation, from the portion of the sample that received excitation radiation, onto at least one optical element which directs the emission radiation along at least one detection path within the housing;
• (c) directing excitation radiation to be incident upon different portions of a sample by relatively moving the beam and the sample;
• (d) for each of a plurality of different portions of the sample, detecting a group of emission signals, with each group comprising signals from a plurality of different wavelength bands, and using at least one detector located within the housing at a location along the at least one detection path;
• (e) for the plurality of groups of emission signals associated with the plurality of different sample portions, distinguishing useful signal groups from unusable signal groups, where useful signal groups are those containing at least one signal having a strength that is greater than a strength of a corresponding background signal by a predefined amount, and wherein each useful signal group is associated with one of the plurality of different sample portions;
• (f) providing both predetermined biohazard indicative signal information and predetermined non-biohazard indicative signal information related to the selected biohazard signature;
• (g) producing a biohazard indicative status for each useful signal group based at least in part upon a comparison of emission signal data for that group with the predetermined biohazard indicative signal information and predetermined non-biohazard indicative signal information;
• and
• (h) forming a biohazard indicative conclusion based, at least in part, on a combination of (1) biohazard indicative signal information for a plurality of useful signal groups, and (2) relative spacings between the portions of the sample associated with the plurality of useful signal groups having biohazard indicative signal information,
• wherein the excitation radiation comprises radiation with wavelength selected from the group consisting of:
• (1) below 300 nanometers, (2) below 275 nm, and (3) below 250 nm,
• wherein the selected biohazard signature is selected from the group consisting of:
• (1) a molecular indicator of a virus,
• (2) a molecular indicator of a non-infecting presence of a virus,
• (3) a molecular indicator of an infecting presence of a virus, and
• (4) a molecular indicator of a past infecting presence of a virus, and
• wherein the selected biohazard is further selected from the group consisting of:
• (1) a corona virus, (2) SARS-CoV-2, (3) an influenza virus (selected from the group consisting of A, B, C, and D), (4) a hemorrhagic fever virus, and (5) Ebola.