Diagnosing and indicating method for non-small cell lung cancer and bioinstrumentation kit

A technology of non-small cell lung cancer and a kit, which is applied in the field of tumor diagnosis and can solve problems that have not been reported yet

Inactive Publication Date: 2015-12-16
崔长友
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, there has been no report on the relationship between CystatinSN and CystatinS in the family of cysteine ​​protease inhibitors and non-small cell lung cancer

Method used

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  • Diagnosing and indicating method for non-small cell lung cancer and bioinstrumentation kit
  • Diagnosing and indicating method for non-small cell lung cancer and bioinstrumentation kit
  • Diagnosing and indicating method for non-small cell lung cancer and bioinstrumentation kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Magnetic bead method specifically enriches CystatinSNmRNA, CystatinSmRNA and RPN1mRNA in urine

[0024] According to the mRNA sequence design of CystatinSN, CystatinS and RPN1 (ribosome binding protein 1), the specific probe (the nucleotide sequence of CystatinSNmRNA such as SEQIDNO.1, the nucleotide sequence of CystatinSmRNA such as SEQIDNO.2, The nucleotide sequence of RPN1mRNA is as SEQIDNO.3): the probe for capturing CystatinSNmRNA is 5'-aaagagcacaactgtttcttctgca(dA)30-3'(SEQIDNO.4), the probe for capturing CystatinSmRNA is 5'-taccaggtctattagaagca(dA)30- 3' (SEQ ID NO.5), the probe for capturing the internal reference gene RPN1 mRNA is 5'-gatgagcttctcattctcaatgtacg(dA)30-3' (SEQ ID NO.6). The above-mentioned specific probes can complementarily bind to olig(dT) of magnetic beads (GE, catalog number: 3815-2103-010150) to obtain magnetic beads bound to specific probes.

[0025] The enrichment steps are as follows: fresh urine and sample transport preservati...

Embodiment 2

[0037] Example 2: Detection of the expression of CystatinSNmRNA and CystatinSmRNA in non-small cell lung cancer tissues

[0038] 30 cases of non-small cell lung cancer and adjacent paired tissue samples were collected from the Department of Oncology, Jiangsu Cancer Hospital, cut to the size of rice grains, stored in RNAlater preservation solution at -80°C, and equilibrated to room temperature before use. Then according to the method of Example 1, the CystatinSNmRNA, CystatinSmRNA and RPN1mRNA of non-small cell lung cancer and adjacent paired tissues were enriched respectively, and then the detection primers of CystatinSNmRNA, CystatinSmRNA detection primers and internal reference gene RPN1mRNA detection primers were used to detect 30 samples of non-small cell lung cancer The relative expression levels of CystatinSNmRNA and CystatinSmRNA in the paired adjacent tissues, the detection system and detection conditions are the same as those in Example 1. The test results are 2 -△△C...

Embodiment 3

[0039] Example 3: Construction of a non-small cell lung cancer detection kit

[0040] 1. Construction of CystatinSN recombinant plasmid

[0041] The non-small cell lung cancer cell line HCC827 was used as the material to extract the total mRNA of the non-small cell lung cancer cell line, and then the extracted mRNA was used as the template to synthesize cDNA, and the primers for constructing the CystatinSN recombinant plasmid were designed according to the sequence of the CystatinSN gene (or CST1 gene), CystatinSN The upstream primer of the recombinant plasmid is 5'-ctggagccccaaggagga-3' (SEQ ID NO.13), the downstream primer is 5'-accagtccagggggtggga-3' (SEQ ID NO.14), and the nucleotides shown in SEQ ID NO.13 and SEQ ID NO.14 are used as primers , the synthesized cDNA was used as a template for PCR amplification. The amplification conditions were: pre-denaturation at 94°C for 5 minutes; 45 cycles of denaturation at 94°C for 30 seconds, annealing at 60°C for 30 seconds, and ex...

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Abstract

The invention discloses a diagnosing and indicating method for the non-small cell lung cancer and a bioinstrumentation kit. Multiple tumor markers are jointly applied to diagnosing and indicating of the non-small cell lung cancer, and the specificity and sensitivity are both remarkably higher than those of diagnosing and indicating the non-small cell lung cancer through a single tumor marker. The invention further provides the bioinstrumentation kit based on multiple markers. The kit is convenient to use, good in specificity, high in sensitivity, capable of being used for diagnosing of the non-small cell lung cancer, treating effect evaluation in the treating process and monitoring of the transferring and relapsing after treatment, and very high in clinical application prospect.

Description

technical field [0001] The invention belongs to the field of tumor diagnosis, and in particular relates to a diagnosis and prediction method and a biological detection kit for non-small cell lung cancer, which are used for auxiliary diagnosis and prediction of non-small cell lung cancer. Background technique [0002] Lung cancer is a disease that seriously threatens human health. With its morbidity and mortality increasing year by year, it has attracted more and more attention from researchers. In 2013, there were 3 million new cases of lung cancer in the world, accounting for 20% of global tumors. Its mortality rate ranks first in male malignant tumors and second in females (after breast cancer). The number of people who died of lung cancer in the world in 2013 Up to 2.6 million people. The current situation of lung cancer in my country is more serious. From 2005 to 2010, the number of lung cancer cases in my country increased by 168,000. If the number of smokers and air p...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/118C12Q2600/158
Inventor 崔长友
Owner 崔长友
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