Method for detecting cancer cell

Patent metadata

Field	Value
Publication number	US-9645154-B2
Application number	US-201113995585-A
Country	US
Kind code	B2
Filing date	Dec 22, 2011
Priority date	Dec 24, 2010
Publication date	May 9, 2017
Grant date	May 9, 2017

Abstract

A method for detecting a cancer cell, including: contacting in vitro a peripheral blood-containing sample including test cells and a labeling substance; allowing the test cells to incorporate the labeling substance to label the test cells; and detecting, as a cancer cell, a cell showing a higher degree of labeling with the labeling substance than that shown by a normal cell in the peripheral blood-containing sample.

First claim

The invention claimed is:

1. A method for testing a peripheral blood-containing sample for cancer cells, the method comprising:
contacting in vitro test cells present in the blood-containing sample with a labeling substance under conditions sufficient for the test cells to react with the labeling substance to form labeled test cells,
wherein the labeling substance is incorporated into the cancer cells and metabolized to protoporphyrin IX, and the following conditions of (i) and (ii) are satisfied:
(i) contacting in vitro the peripheral blood-containing sample comprising the test cells with the labeling substance in the presence of 0.1 to 10 mM of a Mg ion and/or a Ca ion and at least one selected from the group consisting of a reducing agent and an endocytosis-including substance;
(ii) the test cells are allowed to incorporate the labeling substance from 15 minutes to 120 minutes,
wherein the labeling substance is at least one selected from the group consisting of aminolevulinic acid, a salt of aminolevulinic acid, an ester of aminolevulinic acid and a salt of an ester of aminolevulinic acid, and
wherein the method is sufficiently sensitive to detect a cancer cell in the sample when the sample has a cancer cell density of 0.05 cells/mL to 5,000 cells/mL.

2. The method according to claim 1 , wherein the testing includes determining a fluorescence intensity of the test cells labeled with the protoporphyrin IX.

3. The method according to claim 1 , wherein the testing includes determining at least one selected from the group consisting of a mean fluorescence intensity and a total fluorescence intensity of the test cells labeled with the protoporphyrin IX.

4. The method according to claim 1 , wherein the testing is performed on the basis of size of the test cells labeled with the protoporphyrin IX.

5. The method according to claim 1 , wherein the peripheral blood-containing sample is hemolyzed.

6. The method according to claim 1 , wherein the contacting in vitro of the peripheral blood-containing sample with the labeling substance is conducted in the presence of 0.1 to 10 mM of the Mg ion and/or the Ca ion and an endocytosis-inducing substance.

7. The method according to claim 1 , wherein the reducing agent in the contacting in vitro of the peripheral blood-containing sample with the labeling substance is ascorbic acid.

8. The method according to claim 1 , wherein the test cells react with the labeling substance at a temperature of from 1° C. to 42° C.

9. The method according to claim 1 , wherein the method further comprises contacting in vitro the test cells present in the peripheral blood containing sample with a cell-type distinguishing antibody.

10. The method according to claim 9 , wherein the cell type-distinguishing antibody comprises at least one selected from the group consisting of an anti-CD45 antibody and an anti-CD34 antibody.

Assignees

• Takagi Hidenori
• Ito Hiroshi
• Kozuka Masahiro
• Arkray Inc

Inventors

• Takagi Hidenori
• Ito Hiroshi
• Kozuka Masahiro

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• G01N33/575

  for cancer · CPC title

• G01N33/72

  involving blood pigments, e.g. haemoglobin, bilirubin {or other porphyrins; involving occult blood} · CPC title

• G01N33/582 Primary

  with fluorescent label · CPC title

• G01N33/574

  Physics · mapped topic