Method for measuring the concentration of polynucleotides

• US 6,423,490 B1
• Filed: 10/22/1997
• Issued: 07/23/2002
• Est. Priority Date: 10/25/1996
• Status: Expired due to Fees

• Current Assignee: Schott Lithotec AG (Carl-Zeiss-Stiftung)
• Original Assignee: Kyoto Daiichi Kagaku Company Limited
• Inventors: Takama, Toshio
• Primary Examiner(s): FREDMAN, JEFFREY NORMAN
• Application Number:
  US08/955,727
• Time in Patent Office: 1,735 Days
• Field of Search: 435/6, 435/91.2, 435/94, 536/23.1, 536/24.3, 536/24.31, 536/24.32, 536/24.23
• US Class Current: 435/6.14
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 2545/114 involving a quantitation step
• CPC Class Codes: C12Q 2563/113 the label being electroactive, e.g. redox labels
• CPC Class Codes: Y10S 435/912 Absidia

Abstract

A method for measuring the presence of polynucleotide in a sample, comprising the steps of: (1) preparing a sample, an agglutinative agent and an agglutination promoter capable of binding to polynucleotide; (2) mixing said sample, said agent and said promoter; and (3) measuring the degree of agglutination of the agent.

Claims

1. A method for determining whether a target polynucleotide in a sample subjected to polynucleotide amplifying has been amplified, comprising the steps of:

(a) adding to the sample an agglutinative agent and an agglutination promoter capable capable of binding to polynucleotide;

(b) detecting the agglutination of the agglutinative agent; and

(c) determining on the basis of the agglutination whether the target polynucleotide in the sample has been amplified.

2. A method as claimed in claim 1, wherein the polynucleotide is double stranded deoxyribonucleic acid (DNA).

3. A method as claimed in claim 2, wherein the sample is one or more of the samples selected from the group consisting of a sample treated by the polymerase chain reaction (PCR) method, a sample treated by the Strand displacement amplification (SDA) method and a sample treated by Ligase chain reaction (LCR) method.

4. A method as claimed in claim 1, wherein the polynucleotide is ribonucleic acid (RNA).

5. A method as claimed in claim 4, wherein the sample is treated by the Qβ

method with Qβ

-replicase.

6. A method as claimed in claim 1, wherein the polynucleotide is single stranded DNA.

7. A method as claimed in claim 1, wherein the polynucleotide is a complex of RNA and DNA.

8. A method as claimed in claim 1, wherein said promoter is one or more of the promoters selected from the group consisting of 4′

,6- diamidino-2- phenylindole, ethidium bromide, bisbenzimide and acridine orange.

9. A method as claimed in claim 1, wherein said promoter is one or more of the promoters selected from the group consisting of ethidium bromide, thiazole orange, bisbenzimide and acridine orange.

10. A method as claimed in claim 1, wherein said agent is one or more of the agents selected from the group consisting of a silver colloid, a gold colloid, a copper colloid and a latex.

11. A method as claimed in claim 1, wherein step (b) is carried out by visual observation.

12. A method as claimed in claim 1, wherein step (b) is carried out by measuring the absorbance of radiation using a spectrophotometer.