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Detection kit for diagnosis of patient with lung cancer based on multiple genes

A kit and gene technology, applied in the biological field, can solve the problems of low survival rate and achieve high sensitivity

Inactive Publication Date: 2018-07-31
BEIJING EXELLON MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although a series of advances have been made in the diagnosis, surgical treatment and research of lung cancer in recent years, its five-year survival rate has always remained at a low 16%.

Method used

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  • Detection kit for diagnosis of patient with lung cancer based on multiple genes
  • Detection kit for diagnosis of patient with lung cancer based on multiple genes
  • Detection kit for diagnosis of patient with lung cancer based on multiple genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment one: DNA extraction

[0033] DNA extraction reagent consists of lysis buffer, washing buffer and elution buffer, see Table 1; lysis buffer consists of protein denaturant, detergent, pH buffer and nuclease inhibitor; protein denaturant: hydrochloric acid Guanidine (purchased from sigma company); detergent is: Tween20 (purchased from sigma company); pH buffer is: Tris-HCl (purchased from Aladdin company); nuclease inhibitor is: EDTA (purchased from sigma company) .

[0034] Table 1 Reagents used for DNA extraction

[0035]

[0036]

[0037] In this embodiment, a plasma sample (purchased from Beijing 301 Hospital) is taken as an example to extract plasma circulating tumor DNA. The extraction method includes the following steps: (1) Take 1ml of plasma, add the same volume of lysis buffer, and then add proteinase K and CarrierRNA, Make the final concentrations of 100mg / L and 1μg / ml respectively, shake and mix, and incubate at 55°C for 30min; (2) Add 100μl ...

Embodiment 2

[0038] Example 2: Bisulfite treatment of DNA

[0039] Bisulfite treatment of DNA is to use bisulfite reagent to process, bisulfite reagent is composed of bisulfite buffer and protection buffer; bisulfite buffer is sodium bisulfite (purchased from Sigma ) and water mixed liquid; protection buffer is an oxygen radical scavenger hydroquinone (purchased from sigma company) and water mixed liquid. The protein denaturant was guanidine hydrochloride (purchased from sigma); the pH buffer was Tris-HCl (purchased from Aladdin); the nuclease inhibitor was EDTA (purchased from sigma). The reagent formula of this embodiment is shown in Table 2.

[0040] Table 2 Reagents used for bisulfite treatment of DNA

[0041]

[0042] In Example 2, the DNA extracted in Example 1 was used as the treatment object, and bisulfite was used to treat the DNA. The specific methods included: (1) preparing a bisulfite buffer solution, weighing 1 g of sodium bisulfite powder, and adding water to prepare in...

Embodiment 3

[0043] Example 3: Detection of DNA methylation by fluorescent quantitative PCR

[0044] In this example, the detection genes are SHOX2, PTGER4, PCDHGA12, HOXA9 and FOXL2 genes, and the internal reference gene is ACTB. The internal reference primer pair of the internal reference gene ACTB is a primer pair composed of DNA shown in sequence 46 of the sequence listing and the DNA shown in sequence 47 of the sequence listing; the nucleotide sequence of the internal reference probe of the internal reference gene ACTB is shown in sequence 48 of the sequence listing . In this example, the DNA treated with bisulfite in Example 2 is used as a template for PCR amplification.

[0045] The final concentration of the fluorescent quantitative PCR amplification reaction system is composed of: 1 × PCR buffer (purchased from NEB Company), 0.5 mM dNTPs (purchased from NEB Company), 0.5 μM target gene detection primer, 0.2 μM target gene fluorescent probe, 1 μM Target gene blocking primer, 0.3 ...

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Abstract

The invention relates to a detection kit for diagnosis of a patient with lung cancer based on multiple genes. The detection kit comprises a primer probe composition A, a primer probe composition B, aprimer probe composition C, a primer probe composition D and a primer probe composition E, wherein the primer probe composition A comprises a specific primer pair A, a blocking primer A and a probe A;the primer probe composition B comprises a specific primer pair B, a blocking primer B and a probe B; the primer probe composition C comprises a specific primer pair C, a blocking primer C and a probe C; the primer probe composition D comprises a specific primer pair D, a blocking primer D and a probe D; and the primer probe composition E comprises a specific primer pair E, a blocking primer E and a probe E. The kit provided by the invention can be applied to diagnosis of the lung cancer, and provides a novel, rapid, reliable and accurate way for the diagnosis of the lung cancer; and the kitis expected to play an important role in the field of medical detection.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a detection kit for diagnosing lung cancer patients based on multiple genes. Background technique [0002] Lung cancer is a malignant tumor with the highest morbidity and mortality rate in the world, and it is a major disease that threatens human life and health. The World Cancer Report released by the International Agency for Research on Cancer (IARC) of the World Health Organization (IARC) in 2014 shows that the global burden of cancer continues to increase, and lung cancer ranks first among the new common cancers in 2012, accounting for about 1.8 million cases, accounting for 20% of the total number of common cancers. 13%; among the common death cancers, lung cancer also ranks first, about 1.6 million cases, accounting for 19.4% of the total, of which the cases in China account for more than 1 / 3. In addition, my country's 2013 tumor registration annual report shows that the incid...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/166C12Q2600/154
Inventor 李明明索伟克蒲珏
Owner BEIJING EXELLON MEDICAL TECH CO LTD
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