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A kind of lung cancer detection reagent and kit

A technology for detection reagents and kits, which is applied in the field of lung cancer detection reagents and kits, can solve the problems of limitation, patient loss, non-invasive screening of lung cancer, etc., and achieve high specificity and sensitivity, and a wide range of applications

Active Publication Date: 2022-07-12
CREATIVE BIOSCIENCES (GUANGZHOU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

4) Bone scan: It has high sensitivity for detecting bone metastases of lung cancer, but has a certain false positive rate
If an early tumor screening product cannot screen out all or most of the patients, those who miss the test will not be able to receive sufficient risk warnings, thus delaying the timing of treatment, which is a huge loss for patients loss
[0017] Although some tumor markers related to lung cancer have been found in the prior art, the sensitivity and specificity of these tumor markers cannot meet the demand due to the limitation of detection reagents or detection methods for these tumor markers. Therefore, the current There is still a need for further research in the field of screening methods that can actually be applied to lung cancer
However, although non-invasive screening has unique advantages in sampling, it also has some limitations in other aspects, such as adenocarcinoma in lung cancer, which is difficult to cough up through sputum due to exfoliated cells in the deep lung Generally speaking, those skilled in the art would think that this type of lung cancer is not suitable for non-invasive screening
On the other hand, even for other types of lung cancer, the currently reported non-invasive screening methods are difficult to meet the requirements of clinical use
Although relevant research has been progressing for many years, there is still no non-invasive screening method for lung cancer that can be promoted to the clinic

Method used

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  • A kind of lung cancer detection reagent and kit
  • A kind of lung cancer detection reagent and kit
  • A kind of lung cancer detection reagent and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] The inventors screened hundreds of gene markers and nucleic acid fragments, studied the distribution of methylation sites of each gene, and designed primers and probes for real-time fluorescence quantitative methylation-specific polymerase chain reaction (real-time fluorescent quantitative PCR). quantitative methylation-specific PCR, qMSP) detection. Screening is performed in tissue samples, and the β-actin gene is used as an internal reference gene, and the nucleic acid fragment shown in SEQ ID NO: 4 is finally obtained through screening, which has better detection results for lung cancer. The detection effect of SEQ ID NO: 4 fragment and commonly used lung cancer detection gene markers (PCDHGA12, HOXD8) was compared, and each gene detection primer probe was as follows:

[0089] The detection primers and probes for nucleic acid fragments are:

[0090] SEQ ID NO: 1 Primer F: TCGTGTGTCGTCGTTCAGAC

[0091] SEQ ID NO: 2 Primer R: GAAATACCCGCGAAAATACTG

[0092] SEQ ID NO...

Embodiment 2

[0129] Example 2: Detection in sputum samples

[0130] Sample information: A total of 107 sputum samples were tested, including 51 normal control samples, 56 cancer control samples, 20 squamous cell carcinomas, 8 small cell carcinomas, 20 adenocarcinomas, and 56 cancer samples. 1 case of cell carcinoma, 1 case of giant cell carcinoma, and 6 cases of unclassified lung cancer.

[0131] Experimental procedure:

[0132] In this example, the primer probe sequences of the nucleic acid fragments, the primer probe sequences of β-actin, and the DNA modification are the same as those in Example 1.

[0133] a. Collect sputum samples from patients diagnosed with lung cancer and patients with non-lung cancer. After thickening with NaOH, centrifuge to separate cells, wash twice with PBS, and then use Magen DNA extraction kit (HiPureFFPE). DNA Kit, D3126-03) to extract DNA.

[0134] b. The liquid dispensing system is as follows:

[0135] Table 4 dosing system

[0136]

[0137] c. The...

Embodiment 3

[0149] Example 3: Detection in lavage fluid

[0150] Sample information: A total of 176 samples of bronchoalveolar lavage fluid were tested, including 94 samples from the normal control group, 82 samples from the cancer group, 20 samples from the 82 samples from the cancer group, 20 samples from squamous cell carcinoma, 40 samples from adenocarcinoma, and 9 samples from small cell carcinoma. , 13 cases of unspecified type of lung cancer. The primer probes for each gene detection are as follows:

[0151] The detection primers and probes for nucleic acid fragments are:

[0152] SEQ ID NO: 1 Primer F: TCGTGTGTCGTCGTTCAGAC

[0153] SEQ ID NO: 2 Primer R: GAAATACCCGCGAAAATACTG

[0154] SEQ ID NO: 3 Probe P: FAM-AGTTTTACGTTGGAGAAGCGTCGG-BQ1

[0155] The detection primers and probes for β-actin are:

[0156] SEQ ID NO: 6β-actin Primer F: GGAGGTTTAGTAAGTTTTTTGGATT

[0157] SEQ ID NO: 7β-actin primer R: CAATAAAACCTACTCCTCCCTTA

[0158] SEQ ID NO: 8β-actin probe P: Texas Red-TTGT...

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Abstract

The invention belongs to the field of gene detection, and in particular relates to a lung cancer detection reagent and kit. The reagent or kit includes a detection reagent for methylation of a specific nucleic acid fragment and is used to detect the modified sequence of the specific nucleic acid fragment. Tests prove that the reagent of the invention can detect and diagnose lung cancer with high sensitivity and high specificity, and has extremely high clinical application value.

Description

technical field [0001] The invention belongs to the field of gene detection, and more particularly, the invention relates to a lung cancer detection reagent and a kit. Background technique [0002] Lung cancer is a malignant tumor of the lung originating from the bronchial mucosa, glands or alveolar epithelium. According to the pathological type, it can be divided into: 1) small cell lung cancer (SCLC): a special pathological type of lung cancer with obvious tendency to distant metastasis and poor prognosis, but most patients are sensitive to radiotherapy and chemotherapy; 2) Non-small cell lung cancer (NSCLC): other pathological types of lung cancer other than small cell lung cancer, including squamous cell carcinoma, adenocarcinoma, large cell carcinoma, etc. There are some differences in biological behavior and clinical course. According to the location of occurrence, it can be divided into: 1) central lung cancer: lung cancer that grows at or above the bronchial openin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11
CPCC12Q1/6886C12Q1/6851C12Q2600/154C12Q2600/166C12Q2600/112C12Q2531/113C12Q2545/101C12Q2523/125C12Q2563/107
Inventor 吴孝林李仕良吴幽治张志伟陈新周古云娟邹鸿志
Owner CREATIVE BIOSCIENCES (GUANGZHOU) CO LTD