Assay of blood or other biologic samples for target analytes

• US 5,635,362 A
• Filed: 05/23/1994
• Issued: 06/03/1997
• Est. Priority Date: 10/30/1992
• Status: Expired due to Fees

• Current Assignee: QBC Diagnostics, Inc. (Madison Medical Diagnostics LLC)
• Original Assignee: Becton, Dickinson & Co
• Inventors: Wardlaw, Stephen C., Levine, Robert A., Rodriguez, Rodolfo R., Malick, Adrien P., Ozinskas, Alvydas J.
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US08/247,336
• Time in Patent Office: 1,107 Days
• Field of Search: 422/55, 422/58, 435/7.24, 435/287, 435/291, 435/296, 435/810, 435/967, 435/287.1, 435/287.2, 435/288.1, 436/514, 436/518, 436/524, 436/528, 436/531, 436/536, 436/538, 436/541, 436/172, 436/800, 436/805, 436/807, 436/810, 436/824, 356/246, 73/61.43, 73/61.51, 73/61.72, 73/444
• US Class Current: 435/7.24
• CPC Class Codes: B01L 3/5021 Test tubes specially adapted for centrifugation purposes centrifuges B04B5/04
• CPC Class Codes: G01N 33/491 by separating the blood components G01N15/05 takes precedence
• CPC Class Codes: G01N 33/5375 by changing the physical or chemical properties of the medium or immunochemicals, e.g. temperature, density, pH, partitioning
• CPC Class Codes: G01N 33/56972 White blood cells
• CPC Class Codes: G01N 33/585 with a particulate label, e.g. coloured latex
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 435/967 Standards, controls, materials, e.g. validation studies, buffer systems
• CPC Class Codes: Y10S 435/971 Capture of complex after antigen-antibody reaction
• CPC Class Codes: Y10S 435/973 Simultaneous determination of more than one analyte
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick
• CPC Class Codes: Y10S 436/824 Immunological separation techniques
• CPC Class Codes: Y10S 436/829 Liposomes, e.g. encapsulation
• CPC Class Codes: Y10T 436/111666 Utilizing a centrifuge or compartmented rotor

Abstract

A patient'"'"'s health may be diagnosed by centrifuging blood samples in a transparent tube, which tube contains one or more bodies or groups of bodies such as floats, inserts, liposomes, or plastic beads of different densities. Each density-defined body carries analyte-capture binding materials such as antigens or antibodies, which are specific to an epitope, or other specific high affinity binding site on a target analyte which target analyte may be in the blood or other sample being tested; and the level of which analyte is indicative of the patient'"'"'s health. At least one labeled binding material which is also specific to an epitope, or other specific high affinity binding site on the target analyte is added to the sample so as to form labeled binding material/analyte/body complexes in the sample. Upon centrifugation, the complexes will settle out in different areas in the tube according to the respective density of the body or bodies; and the degree of label emission of the complex layers can enable qualitative and/or quantitative analyses of the sample to be made. Unbound labeled binding materials will be separated from the complexed layers by the washing action of ascending or descending components of the sample during the centrifugation step. Unbound labeled binding material will thus not interfere with the analysis.

Claims

1. Paraphernalia for assaying a sample of anticoagulated whole blood for a target analyte, said paraphernalia comprising:

a) a transparent tube for holding the sample;

b) a first generally cylindrical insert capture body in said tube, said capture body being coupled with a binding material which is specific to an epitope or other binding site on the analyte, and said capture body having a specific gravity which is different from the specific gravity of reticulocyte cells in the blood sample;

c) a quantity of label material in said tube for binding to and highlighting the analyte; and

d) a separate band in said tube for forming a label intensity reference for use in quantifying the analyte.

2. The paraphernalia of claim 1 wherein said target analyte is a cellular analyte and said label material is a fluorescent stain which binds to genetic material in the cellular analyte.

3. The paraphernalia of claim 2 wherein said stain is acridine orange.

4. The paraphernalia of claim 2 wherein said separate band is on a second generally cylindrical insert in said tube.

5. The paraphernalia of claim 1 wherein said target analyte is a cellular analyte and said binding material and said label material are specific to CD4 or CD34 epitopes on the cellular analytes.