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Kit for detecting bone metastasis of lung cancer

A kit and bone transfer technology, which can be used in the determination/examination of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., and can solve problems such as difficulty in meeting clinical needs.

Active Publication Date: 2013-01-16
杨祚璋
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Currently, the diagnosis methods for bone metastasis of malignant tumors mainly rely on pathological and imaging examinations, such as biopsy examination, X-ray, CT, MR and ECT examinations, etc., and there is no specificity It is difficult to meet the clinical needs by relying solely on ECT to judge the presence or absence of bone metastases and its progress.

Method used

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  • Kit for detecting bone metastasis of lung cancer
  • Kit for detecting bone metastasis of lung cancer
  • Kit for detecting bone metastasis of lung cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Changes in tumor cell microRNA expression before and after bone metastasis of lung cancer

[0027] 1. Materials and methods

[0028] 1. Materials

[0029] The tumor cells were all from the hospitalized cases of lung cancer in the affiliated hospital from 2004 to 2010. The tumor cells of 30 patients with bone metastasis of lung cancer and the tumor cells of 30 patients with non-metastasis of lung cancer were collected respectively.

[0030] 2. Method

[0031] 2.1 Extraction of total RNA from tumor cells before and after bone metastasis of lung cancer

[0032] According to the instructions of the Trizol kit, RNA was extracted from tumor cells of lung cancer patients with bone metastases and non-metastatic lung cancer patients. The integrity of RNA was verified by gel electrophoresis, and the concentration and purity of RNA were determined with a nucleic acid protein analyzer. The total RNA extraction kit was used by Shanghai Huashun Bioengineering Co., Ltd. t...

Embodiment 2

[0078] Example 2 The expression changes of novell_mir_89 in patients with bone metastasis of lung cancer and normal people

[0079] 1. Experimental materials

[0080] The blood of 50 patients with bone metastases from lung cancer and the blood of 50 normal people were collected to verify whether novel_mir_89 is specifically expressed in patients with bone metastases from lung cancer.

[0081] 2. Experimental methods and results

[0082] 1 Primer design and synthesis

[0083] According to the sequence information of novel_mir_89, four primers were designed. The first one is the reverse transcription primer RT89 with a stem-loop structure, whose sequence is shown in the sequence listing SEQ NO.1; the second is the upstream specific primer F89, whose sequence is shown in the sequence listing SEQ NO.2; the third is the downstream universal primer R89, its sequence is shown in the sequence table SEQNO.3; the fourth is the fluorescent probe primer Flu89 needed in fluorescent quant...

Embodiment 3

[0099] Example 3 Preparation and use of a fluorescent quantitative PCR detection kit for lung cancer bone metastases

[0100] 1. Composition of novel_mir_89qRT-PCR kit

[0101]

[0102] 2. Detection of novel_mir_89qRT-PCR

[0103] 2.1 Preparation of serum small RNA

[0104] The blood of 120 patients with bone metastases from lung cancer and 100 normal people were selected. Centrifuge at 12,000 rpm for 10 minutes, and collect the supernatant.

[0105] 2.2 cDNA synthesis

[0106] The obtained serum was subjected to reverse transcription reaction, and the reverse transcription system was: reverse transcription primer RT89SEQ ID NO.1 (2 μmol / L) 1 μl, blood supernatant 4 μl, dNTP mixture (each 2.5mmol / L) 4 μl, 0.1mol / L LDTT 2 μl, SuperScript RNase H reverse transcriptase (200 U / μl) 2 μl (purchased from Invitrogen Company) The reaction conditions are: 37°C water bath for 60 minutes, 95°C for 3 minutes.

[0107] 2.3qRT-PCR detection

[0108] qRT-PCR was amplified on a NanoDr...

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Abstract

The invention discloses a fluorescent quantitative polymerase chain reaction (PCR) assay kit for bone metastasis of lung cancer. The kit comprises the following components: a specific primer, a specific probe, a standard DNA template and a fluorescent quantitative PCR reaction solution. The invention further discloses a using method of the fluorescent quantitative PCR assay kit for the bone metastasis of the lung cancer. The kit can detect the expression quantity of novel_mir_89 quickly and quantitatively by the kit so as to detect the occurrence of the bone metastasis of the lung cancer. The invention is simple, convenient and quick in operation, stable in result and high in sensitivity and specificity.

Description

technical field [0001] The invention relates to a detection kit, in particular to a fluorescent quantitative PCR detection kit for bone metastasis of lung cancer. Background technique [0002] Currently, more than 1 million patients worldwide are diagnosed with lung cancer every year. Malignant tumors can metastasize to different organs in the body. Bone metastasis is one of the common complications of malignant tumors. Bone tissue is the best organ for distant metastasis of malignant tumors. Many types of malignant tumors can metastasize to bone. Common tumors that cause bone metastasis include breast cancer, prostate cancer, and thyroid cancer. Bone metastases of malignant tumors mostly occur between the ages of 50 and 80, and are rare under the age of 40. Bone metastases can affect the bones of the whole body, and the axial bones (spine, pelvis, etc.) and proximal long bones are the predilection sites for bone metastases. Bone metastases are often multiple, and a few ar...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 杨祚璋谢琳徐磊李国奇
Owner 杨祚璋
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