Method for analyzing a target nucleic acid fragment and a kit for analyzing a target nucleic acid fragment

• US 20050266450A1
• Filed: 04/15/2005
• Published: 12/01/2005
• Est. Priority Date: 06/14/2001
• Status: Abandoned Application

• Current Assignee: Fujifilm Corporation (Fujifilm Holdings Corporation)
• Original Assignee: Fuji Photo Film Co Limited (Fujifilm Holdings Corporation)
• Inventors: Mori, Toshihiro, Iwaki, Yoshihide, Makino, Yoshihiko
• Application Number:
  US11/106,612
• Publication Number:
  US 20050266450A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/42 involving phosphatase
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2521/543 Immobilised enzyme(s)
• CPC Class Codes: C12Q 2533/101 Primer extension
• CPC Class Codes: C12Q 2565/301 Pyrophosphate (PPi)
• CPC Class Codes: C12Q 2565/625 being a nucleic acid test strip device, e.g. dipsticks, strips, tapes, CD plates
• CPC Class Codes: G01N 33/523 the element being adapted for a specific analyte

Abstract

An object of the present invention is to provide a method for analyzing a target nucleic acid fragment which can be simply and swiftly carried out by using a small apparatus, a kit for analyzing a target nucleic acid fragment using the method for analysis, and a dry analytical element for quantifying pyrophosphoric acid. The present invention provides a method for analyzing pyrophosphoric acid generated upon polymerase elongation reaction based on certain nucleotide sequence of a target nucleic acid, a kit for analysis for carrying out the above mentioned method for analysis, and a dry analytical element for quantifying pyrophosphoric acid.

Claims

1. A kit which comprises at least one primer complementary with a part of the target nucleic acid fragment to be analyzed, at least one deoxynucleoside triphosphate, at least one polymerase, and a dry analytical element for quantifying pyrophosphoric acid.

2. A kit which comprises at least one primer complementary with a part of the target nucleic acid fragment to be analyzed, at least one deoxynucleoside triphosphate (dNTP), at least one polymerase, pyrophosphatase, and a dry analytical element for quantifying inorganic phosphorus.

3. A dry analytical element for quantifying pyrophosphoric acid which comprises a reagent for converting pyrophosphoric acid into inorganic phosphorus and a reagent layer containing a group of reagent for carrying out a coloring reaction depending of the amount of inorganic phosphorus.

4. The dry analytical element for quantifying pyrophosphoric acid according to the present invention, which comprises a reagent layer containing xanthosine or inosine, pyrophosphatase, purine nucleoside phosphorylase, xanthine oxidase, peroxidase, and a color developer.