Multiplexed analysis of clinical specimens apparatus and method

• US 6,449,562 B1
• Filed: 08/18/1998
• Issued: 09/10/2002
• Est. Priority Date: 10/10/1996
• Status: Expired due to Term

• Current Assignee: Luminex Corporation (DiaSorin S.p.A.)
• Original Assignee: Luminex Corporation (DiaSorin S.p.A.)
• Inventors: Fulton, Jerrold R., Chandler, Van S., Chandler, Mark B.
• Primary Examiner(s): FREDMAN, JEFFREY NORMAN
• Application Number:
  US09/000,286
• Time in Patent Office: 1,484 Days
• Field of Search: 702/19, 702/20, 435/6, 707/104, 707/1
• US Class Current: 702/19
• CPC Class Codes: C12Q 1/00 Measuring or testing processes involving enzymes, nucleic acids or microorganisms measuring or testing apparatus with condition measuring or sensing means, e.g. colony counters, C12M1/34; Compositions therefor; Processes of preparing such compositions
• CPC Class Codes: G01N 33/537 with separation of immune complex from unbound antigen or antibody
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G16B 25/00 ICT specially adapted for hybridisation; ICT specially adapted for gene or protein expression
• CPC Class Codes: G16B 25/20 Polymerase chain reaction [PCR]; Primer or probe design; Probe optimisation
• CPC Class Codes: G16B 40/00 ICT specially adapted for biostatistics; ICT specially adapted for bioinformatics-related machine learning or data mining, e.g. knowledge discovery or pattern finding
• CPC Class Codes: G16B 50/00 ICT programming tools or database systems specially adapted for bioinformatics
• CPC Class Codes: G16B 50/10 Ontologies; Annotations
• CPC Class Codes: Y10S 707/99931 Database or file accessing

Abstract

A method for the multiplexed diagnostic and genetic analysis of enzymes, DNA fragments, antibodies, and other biomolecules comprises the steps of constructing an appropriately labeled beadset, exposing the beadset to a clinical sample, and analyzing the combined sample/beadset by flow cytometry is disclosed. Flow cytometric measurements are used to classify, in real-time, beads within an exposed beadset and textual explanations, based on the accumulated data obtained during real-time analysis, are generated for the user. The inventive technology enables the simultaneous, and automated, detection and interpretation of multiple biomolecules or DNA sequences in real-time while also reducing the cost of performing diagnostic and genetic assays.

Claims

1. A computer readable medium storing an assay database for processing a pooled population of subsets of particles during flow analysis, said assay database comprising:

(a) a discriminant function table encoding a decision tree for classifying in real time each particle according to its subset based at least in part on at least one classification parameter including at least one respective fluorescence emission intensity, and (b) an assay definition table encoding a decision tree for defining an assay by identifying a respective subset and as least one baseline assay measurement value for interpreting an assay result.

2. The computer readable medium according to claim 1, wherein said discriminant function table includes, for each subset, at least one of high and low classification parameter values.

4. The computer readable medium according to claim 1, wherein said assay database further includes an interpretation table including at least one textual message for each assay result based on the at least one baseline assay measurement value.

5. The computer readable medium according to claim 1, wherein said assay database further includes a results table encoding, for each subset, at least one of a total number of particles classified per subset, a running sum of collected assay measurement values, a total number of particles having collected assay measurement values less than the at least one baseline assay measurement value, and a total number of particles having collected assay measurement values at least as great as the at least one baseline assay measurement value.

3. A machine readable assay database, stored in a storage device, for the processing of flow-cytometric measurement data comprising:

(a) an assay definition table, said assay definition table encoding (1) one or more measurement subset token identifiers, (2) for each subset token identifier, one or more baseline measurement parameter values, and (3) for each subset token identifier, an interpretation test-type token;

(b) a discriminant function table, said discriminant function table encoding a real-time classification decision tree based on one or more classification measurement parameters, said one or more classification measurement parameters encoded in said flow-cytometric measurement data;

(c) an interpretation table, said interpretation table encoding textual assay outcome-description information; and

(d) a results table, said results table capable of encoding statistical accumulation of real-time flow-cytometric measurement data.

6. A computer readable medium storing an assay database for processing a pooled population of subsets of particles during flow analysis, said assay database comprising:

(a) a discriminant function table encoding a decision tree for classifying in one step each particle according to its subset based at least in part on at least one classification parameter including at least one respective fluorescence emission intensity, and (b) an assay definition table encoding a decision tree for defining an assay by identifying a respective subset and as least one baseline assay measurement value for interpreting an assay result.

7. A computer readable medium storing an assay database for processing a pooled population of subsets of particles during flow analysis, said assay database comprising:

(a) a discriminant function table encoding a decision tree for classifying each particle according to its subset based at least in part on at least one classification parameter, and (b) an assay definition table encoding a decision tree for defining an assay based at least in part on at least one baseline measurement parameter value.

8. The computer readable medium of claim 7 in which said classification parameter includes at least one fluorescence emission intensity.

9. The computer readable medium of claim 7 in which said at least one classification parameter includes at least two fluorescence emission intensities.

10. The computer readable medium of claim 7 in which said baseline measurement arameter value is used for interpreting an assay result.

11. The computer readable medium of claim 7 in which said assay definition table further encodes at least one measurement subset token identifiers.

12. The computer readable medium of claim 7 in which said assay database further comprises an interpretation table.

13. The computer readable medium of claim 7 in which said assay database further comprises a results table.