Lung cancer early screening kit

A kit and lung cancer technology, applied in the field of molecular biology, can solve the problems of lack of specific early screening molecular markers, limit early screening of lung cancer, and low signal sensitivity, so as to avoid false negative and false positive results and improve Cure and Survival Rates, Effects of Improving Specificity and Accuracy

Inactive Publication Date: 2017-12-15
SUREXAM BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are also mRNA early screening technologies for lung cancer-related tumor genes, such as RT-PCR, real-time PCR and fluorescence in situ hybridization, etc., but the current detection is mainly based on the principle of PCR reaction, which has defects such as easy contamination and high false positive rate. However, the traditional RNA fluorescence in situ hybridization itself has the disadvantage of low signal sensitivity.
In addition, the lack of specific early screening molecular markers is also an important factor limiting the early screening of lung cancer

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] There are two types of lung cancer early screening kits described in this example, with labeled probes and without labeled probes.

[0027] Among them, the lung cancer early screening kit A with labeled probes mainly includes:

[0028] 1. Capture probe

[0029] The capture probe consists of three parts, the 5' end to the 3' end are sequentially the P1 sequence that can be complementary to the mRNA of the corresponding target gene, the spacer sequence, and the P2 sequence that is complementary to the corresponding amplification probe P3 sequence, Target genes of the same category have the same P2 sequence in their capture probes. The spacer is used to space the capture probe P2 sequence from the target mRNA, and by setting a spacer sequence of appropriate length inside the probe, it can reduce steric hindrance, improve the efficiency of the hybridization reaction and the specificity of the hybridization reaction . The spacer arm of the capture probe of the present inv...

Embodiment 2

[0052] Example 2 Using kit A in Example 1 to detect circulating tumor cells in peripheral blood of patients with lung cancer

[0053] The formula of described various solutions is as follows:

[0054]

[0055] The probe mixture, amplification mixture, and chromogenic mixture in this example all use all the probes in the corresponding gene list of the lung cancer early screening kit A with labeled probes in Example 1.

[0056] 1. Sample pretreatment, filter the screening cells onto the filter membrane

[0057] 1. Preserve the blood sample in the sample preservation tube with preservation solution, centrifuge at 600×g for 5 minutes, discard the supernatant, and remove the red blood cells.

[0058] 2. Add 4mL PBS and 1mL fixative, vortex to mix, and let stand at room temperature for 8min.

[0059] 3. Sample filtration: transfer the liquid in the sample storage tube to the filter, turn on the vacuum filtration pump to drain the liquid; add 4mL PBS to the storage tube, wash th...

Embodiment 3

[0108] The selection of embodiment 3 target detection gene quantity and kind

[0109] 1. Design of kit preparation (selection of the number and types of target detection genes)

[0110] The lung cancer screening genes of the kit of the present invention are selected from: TTF-1, LUNX, NSE, EGFR, P63, CEA, and make corresponding selections according to the experimental group. Non-blood-derived cell-related genes are used: CK7, CK19, hTERT, VIM, PIM -1 and Survivin; excluded genes used: CD45, CD34, CD105, CD144, CD146 and VWF.

[0111] For the selection of lung cancer screening genes, refer to experimental groups 1-4, select one, two, three and six target genes respectively, and compare their detection effects, and use all target genes for non-blood-derived cell-related genes and exclusion genes , the specific design is shown in Table 7.

[0112] The compositions and quantities of the capture probes, amplification probes and labeling probes, detection methods, etc. of each gro...

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Abstract

The invention relates to a lung cancer early screening kit. The kit comprises capture probes, amplification probes and labeled probes for detection of target gene mRNAs. The target genes comprise lung cancer screening genes and non-blood cell related genes. The lung cancer screening genes comprise at least two of TTF-1, LUNX, NSE, EGFR, P63 and CEA. The non-blood cell related genes comprise at least two of CK7, CK19, hTERT, VIM, PIM-1 and Survivin. Through screening of target genes, detection of a single target gene is realized and simultaneous detection of multiple target genes is realized so that lung cancer early screening cells can be completely detected, leukocytes, hematopoietic stem cells and endothelial cells in blood are distinguished from the screening cells, false negative and false positive results are avoided and the accuracy of detection is further improved.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a lung cancer early screening kit. Background technique [0002] Lung cancer is a malignant tumor originating from the epithelium of the bronchial mucosa, also known as bronchial lung cancer. Its morbidity and mortality are the fastest growing among malignant tumors, and it is one of the most threatening malignant tumors to human health and life. In the past 50 years, many countries have reported that the morbidity and mortality of lung cancer have increased significantly. The morbidity and mortality of lung cancer in men account for the first of all malignant tumors, and the incidence and mortality of lung cancer occupy the second place in women. . The early diagnosis rate of lung cancer is low. Patients with stage 0 often have no symptoms, and less than 0.6% of the total number of lung cancer patients are diagnosed at the time of diagnosi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/16
Inventor 刘苏燕吴诗扬董艳
Owner SUREXAM BIO TECH
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