Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for diagnosis of cancer and monitoring of cancer treatments

A cancer and liver cancer technology, applied in the field of cancer diagnosis and cancer treatment monitoring, can solve the problems of lack of specificity and hindering the clinical application of cancer diagnosis

Inactive Publication Date: 2012-03-07
奥里迪斯生物标记有限责任公司
View PDF2 Cites 16 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, due to the lack of specificity, the level of circular DNA is not a useful clinical marker for cancer diagnosis, as too many confounding biological and physiological processes seem to affect the level of circular DNA, thereby hindering its clinical application in cancer diagnosis

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for diagnosis of cancer and monitoring of cancer treatments
  • Method for diagnosis of cancer and monitoring of cancer treatments
  • Method for diagnosis of cancer and monitoring of cancer treatments

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Embodiment 1: Method of sample analysis and DNA fragment pattern selection

[0071] In general, sample analysis and fragment mode selection are performed in the following five steps. Step (1) isolation of circular DNA, step (2) quantification of isolated DNA, step (3) quantification of DNA fragments by real-time PCR, step (4) data evaluation and fragmentation pattern selection and finally step (5) DNA fragmentation index ( DFI) calculation.

[0072] Step (1): Isolation of circular DNA

[0073] According to the manufacturer's instructions (Qiagen kit manual, third edition, March 2007, pages 23-25), the circular DNA was extracted from 1 to 2.5ml of frozen (-20°C) or fresh serum samples. Frozen plasma samples (2- 2.5ml).

[0074] Step (2): Quantification of isolated DNA

[0075] The isolated DNA (present in a total volume of up to 20 μl) was quantified using the PicoGreen assay (Cat. No. P11496, Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructi...

Embodiment 2

[0120] Example 2: Recurrence Diagnosis of Metastatic Breast Cancer by DNA Fragmentation Analysis

[0121] DNA fragmentation patterns were recorded for healthy donors (n=17) and a collection of metastatic breast cancer patients (n=10) at different clinical stages (Example 1). LINE1 fragments of indicated bp length (see Table 2) were analyzed and normalized and plotted (see Figure 1B ). DNA fragmentation patterns of pooled breast cancer sera (see Figure 1A ) indicates elevated levels of DNA fragments ranging from 200 to 400 bp compared to healthy controls.

[0122] According to formula [2] (using LINE1-B, LINE1-C, LINE1-E and LINE1-F, see Table 2) and formula [4] (using LINE1-B, LINE1-C, LINE1-D, LINE1-E and LINE1-F, see Table 2), the DNA Fragmentation Index (DFI) was calculated for each patient / collection.

[0123] Mean DFI for healthy controls compared to 0.64 for the breast cancer set 4 (Equation [2]) was 0.02 ± 0.01,; thus suggesting a potential cutoff of approximately...

Embodiment 3

[0134] Example 3: Diagnosis of Hepatocellular Carcinoma by DNA Fragmentation Analysis

[0135] DNA fragmentation patterns were recorded as described in Example 1 for cirrhotic patients (at-risk patients) and HCC patients. LINE1 fragments of the indicated sizes (see Table 2) were analyzed and normalized and plotted (see figure 2 ). figure 2 showed significant differences in DNA fragmentation pattern profiles in HCC patients compared with at-risk patients with cirrhosis. Therefore, a diagnosis of HCC can be obtained by recording the DNA fragmentation pattern in the indicated range of 148 to 783 bp but at least in the range of 148 to 463 bp and comparing it with DNA fragmentation in patients with cirrhosis and / or chronic hepatitis C Schema comparison. The DNA fragmentation pattern differed most significantly between HCC patients and controls between 204 and 463 bp.

[0136] DFI is calculated according to formulas [1], [2] and [4.1]. To optimize the LINE1 model, the differe...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present invention relates to a method for cancer diagnosis and for monitoring cancer treatments based on the analysis of the DNA fragmentation pattern of repetitive elements (preferably LINE1) or multi copy genes (preferably U1 RNA) identified in body fluid samples isolated from cancer patients.

Description

technical field [0001] The present invention relates to a method for cancer diagnosis and monitoring of cancer therapy based on circular DNA analysis, in particular based on the analysis of specific DNA fragmentation patterns of repetitive elements or multi-copy genes identified in bodily fluid samples. Background technique [0002] Cell-free genomic DNA, called circular DNA, is present in low concentrations (0.2 to 200 ng / ml) in serum, plasma and other body fluids (ie urine, ascites, etc.) and is highly degraded (Wang et al., 2003, Cancer Res., 63 (14):3966-8). In the successful detection of circular DNA from tumor patients (Sidransky, D. et al., 1991; Science, 252(5006): 706-9; Kimura, H., et al., 2007, Br.J. Cancer, 97(6): 778-84) in the case of microsatellite instability and tumor-specific mutations (e.g. p53, K-ras, EGFR), genomic DNA (circular tumor DNA) derived from tumor tissue can be observed as cancer patient circular Components of DNA-like DNA (Sozzi, G. et al.,...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/106C12Q2600/156
Inventor M·奥特M·斯瓦茨
Owner 奥里迪斯生物标记有限责任公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products