IMPROVED METHOD FOR PROVIDING INFORMATION ON BREAST CANCER AND DIAGNOSTIC KIT THEREFOR

• US 20200131579A1
• Filed: 02/28/2014
• Published: 04/30/2020
• Est. Priority Date: 02/25/2014
• Status: Abandoned Application

• Current Assignee: Industry-Academic Cooperation Foundation Yonsei University, Optipharm Co., Ltd.
• Original Assignee: Industry-Academic Cooperation Foundation Yonsei University, Optipharm Co., Ltd.
• Inventors: LEE, Hye Young, WANG, Hye Young, KIM, Yeun
• Application Number:
  US15/121,377
• Publication Number:
  US 20200131579A1
• Time in Patent Office: Days
• CPC Class Codes: C12N 15/1096 cDNA Synthesis; Subtracted cDNA library construction, e.g. RT, RT-PCR
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2561/101 Taqman
• CPC Class Codes: C12Q 2561/113 Real time assay
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: C12Q 2600/16 Primer sets for multiplex assays

Abstract

The present disclosure relates to an improved method for providing information on breast cancer and a diagnostic kit therefore, and more specifically, to an improved kit for diagnosing breast cancer by using a one-tube nested PCR and a diagnostic method therefor.

Claims

1. A method for providing information for diagnosis of breast cancer, the method comprising:

a) isolating a total RNA from cells obtained from the blood of a cancer suspected patient;

b) synthesizing cDNA from the isolated total RNA;

c) performing a realtime-PCR of the synthesized cDNA by using one or more primer sets and probes selected from the group consisting of a primer set and a probe capable of amplifying a human epidermal growth factor receptor 2 HER2 and a primer set and a probe capable of amplifying glyceraldehyde-3-phosphate dehydrogenase (GAPDH); and

d) comparing the amplified level with an expressed level in a normal person;

wherein the primer set capable of amplifying the HER2 is a primer set selected from the group consisting of primer sets as set forth in SEQ ID NOS. 1 and 2, SEQ ID NOS. 3 and 4, SEQ ID NOS. 6 and 7, and SEQ ID NOS. 8 and 9 or a mixture of these primer sets and the probe is one or more of probes as set forth in SEQ ID NO. 5, SEQ ID NO. 10, and SEQ ID NO. 11.

2. The method for providing information for diagnosis of breast cancer of claim 1, wherein the comparing of the amplified level with the amplified level in the normal person is performed by a standard or cut-off value.

3. The method for providing information for diagnosis of breast cancer of claim 1, wherein the primer set capable of amplifying the GAPDH is set forth in SEQ ID NOS. 12 and 13 and the probe has a base sequence as set forth in SEQ ID NO. 14.

4. A composition for diagnosing breast cancer, the composition comprising:

a primer sets capable of amplifying a HER2 which is selected from the group consisting of primer sets as set forth in SEQ ID NOS. 1 and 2, SEQ ID NOS. 3 and 4, SEQ ID NOS. 6 and 7, and SEQ ID NOS. 8 and 9 or a mixture of these primer sets; and

a primer set and a probe capable of amplifying GAPDH, as active ingredients.

5. The composition for diagnosing breast cancer of claim 4, wherein the primer set capable of amplifying the GAPDH is set forth in SEQ ID NOS. 12 and 13 and the probe has a base sequence as set forth in SEQ ID NO.14.

8. The composition for diagnosing breast cancer of claim 4, wherein a 5′

-terminal of the probe is marked with a fluorescent material.

9. The composition for diagnosing breast cancer of claim 5, wherein a 5′

-terminal of the probe is marked with a fluorescent material.

10. A kit for diagnosing breast cancer, the kit comprising the composition of claim 4.

11. A kit for diagnosing breast cancer, the kit comprising the composition of claim 5.

6. (canceled)

7. (canceled)