Method for identifying miRNA in blood serum of patient with lung cancer by Solexa technology

A technology of microRNA and patient serum, which is applied in the field of biomedicine, can solve the problems of poor stability and repeatability of information quality, and the inability to realize quantitative detection, etc., and achieve the effects of easy storage, convenient material collection, and low detection cost

Inactive Publication Date: 2008-11-19
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In fact, one of the prominent weaknesses of the gene chip is that the stability and repeatability of its information quality are relatively poor, and quantitative detection cannot be realized.

Method used

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  • Method for identifying miRNA in blood serum of patient with lung cancer by Solexa technology
  • Method for identifying miRNA in blood serum of patient with lung cancer by Solexa technology
  • Method for identifying miRNA in blood serum of patient with lung cancer by Solexa technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Solexa sequencing technology was used to sequence the serum samples of lung cancer patients and analyze the expression levels of each microRNA. The specific steps are:

[0037] (1) Collect serum samples from lung cancer patients.

[0038] (2) Trizol method (Invitrogen Company) extracted total RNA from 100ml of mixed serum of the above patients, concentrated RNA by isopropanol precipitation, and quantified it with a spectrophotometer (100ml serum can usually enrich about 10-20μg of total RNA).

[0039] (3) Apply about 10-20 μg of serum total RNA from lung cancer patients, and use Solexa sequencing technology to detect and analyze RNA samples. The specific plan is:

[0040] a. Apply PAGE electrophoresis to purify and obtain RNA molecules with a length less than 30nt. An adapter is added to both ends of the RNA molecule.

[0041] b. While using the Solexa chip to immobilize one end of the RNA adapter molecule, since the surface of the chip has a single-strand...

Embodiment 2

[0065] The changes of microRNA in the serum of patients with lung cancer compared with normal physiological state in embodiment 2

[0066] Normal human serum microRNA was sequenced by Solexa technology, and the results are shown in Table 3.

[0067] Table 3 Solexa sequencing results of normal human serum

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[0074] 199 microRNAs (see Table 2) were detected in the serum of lung cancer patients, and 167 microRNAs (see Table 3) were detected in normal human serum. We believe that when the change factor of the microRNA is greater than 5, and the difference in absolute value is greater than 20 copy numbers, then the microRNA is significantly changed, otherwise it is regarded as unchanged. According to this rule, we compared the microRNAs in the serum of lung cancer patients and normal people, and found that 63 microRNAs showed an upward trend, 34 showed a downward trend, and 149 remained unchanged. We analyzed ...

Embodiment 3

[0075] Embodiment 3 is used for the making of the serum microRNA test kit of diagnosing lung cancer

[0076] The production process and operation process of the microRNA kit for diagnosing and predicting lung cancer are based on Solexa sequencing technology, combined with quantitative and semi-quantitative PCR technology.

[0077] Through the Solexa sequencing method, the expression profiles of serum microRNAs in normal people and lung cancer patients are first determined, and then a series of serum microRNAs with large expression levels and differences in lung cancer and normal physiological conditions are screened to predict whether lung cancer occurs and diagnose indicator of disease severity. Table 4 shows that compared with normal human serum, the significantly changed microRNAs in the serum of lung cancer patients can screen out a batch of serum microRNAs as molecular markers for the diagnosis of lung cancer.

[0078] Table 4: MicroRNAs changed in lung cancer serum comp...

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Abstract

A method for identifying micro ribonucleic acid (RNA) in the blood serum of a lung cancer patient through Solexa technology belongs to the biological medicine technical field, in particular relating to a Solexa technology for measuring the expression of micro RNA in the blood serum of a lung cancer patient as well as an application of the Solexa technology in clinic diagnosis and therapy such as early diagnosis of lung cancer, curative effect appraisement and drug effect appraisement, etc. The Solexa technology comprises the following steps that: a Solexa sequencing method is used to measure the content of micro RNA in the blood serum of a lung cancer testee; the appraisement of the physiological and / or pathological status of the testee is to carry out lung cancer diagnosis of the testee; and the physiological and / or pathological status of the testee is tested after the testee receives a to-be-tested object. The Solexa sequencing method has the advantages of wide detection pedigree, high sensitivity, low detection cost, convenient material obtaining and easily stored samples, etc.; moreover, the method can be widely used in related operations including lung cancer screening, etc. so as to improve the conditions of low specificity and low sensitivity caused by individual difference which is difficult to be overcome by a single marker; therefore, the method can substantially increase the clinic detection rate of lung cancer, and is an effective way to carry out early diagnosis of lung cancer.

Description

1. Technical field [0001] The invention belongs to the technical field of biomedicine. It specifically relates to a Solexa technology for measuring the expression of microRNA in serum of lung cancer patients, and the application of Solexa technology in clinical diagnosis and treatment of early diagnosis, curative effect evaluation and drug effect evaluation of lung cancer. 2. Background technology [0002] Lung cancer is the most common malignant tumor with the highest morbidity and mortality rate and poor treatment effect. Due to the lack of specific clinical symptoms and manifestations in the early stage of lung cancer, most of the patients are already in the advanced stage when they are waiting for treatment, and the clinical treatment effect and median survival time are unsatisfactory. The overall survival rate of lung cancer has not improved significantly in the past 20 years, and the current 5-year survival rate after surgery is only 14%. Early detection and diagnosi...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 陈江宁王可慧高嵩张峻峰张辰宇蔡星殷媛郭继刚陈熹王进曾柯
Owner NANJING UNIV
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