Patent ID: 8173359

Claim:
A method for live/dead assay for bacterial spores in a specimen, the method comprising: providing a first positively charged multidentate ligand lanthanide complex in a predetermined amount of a first sample from the specimen of live and dead bacterial spores; releasing Dipicolinic Acid (DPA) from the live bacterial spores by germination in the first sample; combining the positively charged multidentate ligand lanthanide complex with the DPA released from the bacterial spores in the predetermined amount of the first sample; exciting the combined positively charged multidentate ligand lanthanide complex and DPA released from the live bacterial spores to generate a first luminescence characteristic of the combined positively charged multidentate ligand lanthanide complex and DPA to detect a number of live bacterial spores in the first sample; providing a second positively charged multidentate ligand lanthanide complex in predetermined amount of a second sample from the specimen of live and dead bacterial spores; releasing DPA from the second sample of bacterial spores by autoclaving, sonication or microwaving; combining the positively charged multidentate ligand lanthanide complex with the DPA released from the bacterial spores in the second predetermined amount of the second sample; exciting the combined positively charged multidentate ligand lanthanide complex and DPA released from the bacterial spores to generate a second luminescence characteristic of the combined positively charged multidentate ligand lanthanide complex and DPA to detect a number of total bacterial spores in the second sample, said exciting comprising radiating the combined positively charged multidentate ligand lanthanide complex and DPA with ultraviolet light; determining a number of dead bacterial spores by subtracting the number of live bacterial spores from the number of total bacterial spores; and generating a ratio of the DPA released from the live bacterial spores in the first sample to the DPA released from the dead bacterial spores in the second sample to obtain a live:dead ratio for the specimen.