Application of fingerprint spectrum consisting of small RNA to diagnosis and treatment of human malignant pleural effusion
A malignant pleural effusion, pleural effusion technology, applied in bioengineering and detection, biomedical fields, can solve the problem that pleural effusion is difficult to distinguish between benign and malignant
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[0140] The preparation of the miRNA chip can adopt conventional manufacturing methods of biochips known in the art. For example, if the solid phase carrier is a modified glass slide or silicon wafer, and the 5' end of the probe contains amino-modified poly dT strings, the oligonucleotide probe can be formulated into a solution, and then spotted with a spotting instrument. The miRNA chip of the present invention can be obtained by arranging in a predetermined sequence or array on a modified glass slide or a silicon chip, and then fixing it by standing overnight. If the nucleic acid does not contain amino modification, its preparation method can also refer to: "Gene Diagnosis Technology-Non-Radioactive Operation Manual" edited by Wang Shenwu; J.L.erisi, V.R.Iyer, P.O.BROWN. Exploring the metabolic and genetic control of gene expression on a genomic scale. Science, 1997; 278:680 and Ma Liren, edited by Jiang Zhonghua. Biochip. Beijing: Chemical Industry Press, 2000, 1-130.
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Embodiment 1
[0167] Sample collection and information analysis
[0168] The samples were approved by the Ethics Committee of Shanghai Zhongshan Hospital, and were obtained in accordance with the method of collecting pleural effusion in the Pathology Department of Shanghai Zhongshan Hospital under the condition that the patients signed the informed consent. A total of 236 pleural effusion samples were obtained from patient chests and stored at -80°C. The samples were divided into two parts, one part of the samples was used as the training set for establishing the analysis model (77 cases), and the other part of the samples was used as the double-blind verification set for double-blind verification (159 cases). The sample information is shown in Table 1. The ratio of male to female in the sample is about 1.5:1, and the average age of the sample is about 61.6 years old.
[0169] Table 1 Sample information
[0170]
Embodiment 2
[0172] Total RNA Extraction
[0173] The above fresh pleural effusion was collected and pretreated by centrifugation, and the sample sediment and supernatant were placed in a -80°C refrigerator for freezing.
[0174] Total RNA was extracted from pleural effusion samples according to the instructions of miRNeasy MiniKit (Qiagen, 217004). The specific operation steps were carried out according to Xiangqiong Company's method for extracting total RNA from pleural fluid.
[0175] The quality was detected by electrophoresis, the OD260nm and OD280nm were measured by ultraviolet spectrophotometry, and the RNA concentration was calculated. Store at -80°C.
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