Claims Analysis
Total Claims: 7
Claim 1
Independent Claim
136 words
Method of analyzing a urine specimen, characterized in that for the rapid and simultaneous counting of both viable bacteria and leucocytes (white blood cells or WBC), it consists of: (1) simultaneous labeling of said bacteria and said white blood cells (WBC) present in said urine specimen, with a fluorescent viability marker of bacteria, said marker tagging and accumulating in both bacteria and white blood cells, under the same conditions and leading to different fluorescent elements; (2) automatically counting all the fluorescent elements by photoelectric detection of the fluorescent marker; and (3) simultaneously classifying said fluorescent elements in fluorescent viable bacteria and in fluorescent white blood cells, in view to obtain in one part the total number of bacteria and on the other part, the total number of white blood cells, on the basis of their size.
Claim 2
Dependent Claim
38 words
Method according to claim 1, characterized in that said viability marker of bacteria of step (1) is selected in the group consisting of products made fluorescent by an intracelluar enzyme, for instance a non-specific esterase or a phosphatase.
Claim 3
Dependent Claim
40 words
Method according to claim 1, characterized in that step (1) comprises, when said viability marker of bacteria is made fluorescent by an intracellular non-specific esterase, incubating a urine specimen with a solution of carboxyfluorescein diacetate at 0.1-0.5%, at neutral pH.
Claim 4
Dependent Claim
36 words
Method according to claim 3, characterized in that said incubation step includes a first incubation of 5-15 minutes, preferably 10 minutes at 37°C followed by a second incubation of 5-15 minutes, preferably 10 minutes at 4°C.
Claim 5
Dependent Claim
29 words
Method according to claim 1, characterized in that, when said labeling is performed on a nonsedimented sample of urine, steps (2) and (3) are carried out by flow cytometry.
Claim 6
Dependent Claim
181 words
Method of analyzing a urine specimen, characterized in that for the rapid and simultaneous counting of both viable bacteria and leucocytes (white blood cells or WBC), it consists of: (1) quantitative concentration of the urine sample by filtering a known quantity of said sample through a filter with pore size small enough to retain bacteria and white blood cells, (2) simultaneous labeling of the elements retained on said filter, with a fluorescent viability marker of bacteria, said marker tagging and accumulating in both bacteria and white blood cells, under the same conditions and leading to different fluorescent elements, (3) counting and classifying fluorescent elements on the whole filter by means of an appropriate scanning cytometer, such as to obtain on one part the number of fluorescent bacteria and on the other part, the number of fluorescent leucocytes, on the basis of their size and (4) calculating the concentration in bacteria and in leucocytes present in the initial urine sample by dividing the obtained counts by the volume of urine filtered, to express them in terms of concentration in the original sample.
Claim 7
Dependent Claim
40 words
Method according to any one of claims 1 to 6, characterized in that the size of said fluorescent elements is evaluated either by measuring the fluorescent intensity or the scattering light intensity or aperture impedance of the different fluorescent elements.