Method of assaying specimen substance by controlling dose of chemiluminescence

• US 5,952,238 A
• Filed: 10/08/1997
• Issued: 09/14/1999
• Est. Priority Date: 02/02/1995
• Status: Expired due to Term

• Current Assignee: Chugai Seiyaku Kabushiki Kaisha (Roche Holding AG)
• Original Assignee: Chugai Seiyaku Kabushiki Kaisha (Roche Holding AG)
• Inventors: Tsuji, Yasuhiro, Kamisango, Keiichi, Otsuka, Mitsuo
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): CARRILLO, BIBI SHARIDAN
• Application Number:
  US08/875,592
• Time in Patent Office: 706 Days
• Field of Search: 73/61.48, 73/64.56, 250/361 C, 250/362, 250/364, 422/52, 422/68.1, 422/82.05, 436/27, 436/56, 436/171, 436/172, 435/6, 435/91.2
• US Class Current: 436/172
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2527/125 Specific component of sample, medium or buffer
• CPC Class Codes: C12Q 2563/103 luminescence
• CPC Class Codes: C12Q 2565/107 Alteration in the property of hybridised versus free label oligonucleotides
• CPC Class Codes: G01N 21/76 Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/825 Pretreatment for removal of interfering factors from sample
• CPC Class Codes: Y10T 436/13 Tracers or tags

Abstract

A method for assay of an analyte by use of a material labeled with a chemiluminescent substance, which comprises adding a quencher and/or decreasing the specific activity of a chemiluminescent substance labeled probe, thereby decreasing the quantity of chemiluminescence.

Claims

1. A method for assay of an analyte which comprises the steps of:

(1) providing a first probe that specifically binds to said analyte wherein said first probe is labeled with a chemiluminescent substance;

(2) adding an unlabeled, second probe that specifically binds to said analyte;

(3) contacting said first labeled probe and said unlabeled second probe with said analyte;

(4) quantitatively measuring a chemiluminescent signal, wherein said unlabeled second probe decreases the chemiluminescent signal by decreasing a specific activity of the chemiluminescent substance of said first labeled probe;

(5) adding a quencher to said analyte before said measuring step to decrease said chemiluminescent signal and decrease background noise levels of said chemiluminescent signal; and

(6) correlating said chemiluminescent signal to the presence of said analyte.

2. The method of claim 1, wherein said quencher is at least one member selected from the group consisting of color matter and india ink.

3. The method of claim 1, wherein said unlabeled probe is added in an amount of from 10 to 10³ parts per one part of the labeled probe.

4. The method of claim 2, wherein said color matter is selected from the group consisting of Crystal Violet, Bromophenol Blue, carminic acid, Chlorophenol Red, hematoxylin, Bromophenol Purple, Bromophenol Red, rosolic acid, Phenol Red, Cresol Red, and Methacresol Red.