Fingerprint spectrum consisting of small RNAs and application thereof to diagnosis of bladder cancer
A technology of bladder cancer and collection, applied in the fields of biomedicine, bioengineering and detection, can solve the problems of low sensitivity, inability to diagnose patients' compliance in early stage, etc.
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[0109] 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
[0132] Sample collection and information analysis
[0133] The samples in this study were frozen tissue samples collected from Zhongshan Hospital Affiliated to Fudan University between March 2016 and November 2016. All samples were collected with personal consent and approved by the hospital's ethics committee, and pathological diagnosis results were confirmed by more than two pathologists. A total of 59 samples, including 30 samples of bladder cancer, 29 samples of paracancerous (normal) tissue, aged from 46 to 90 years (average 71.1 years). Samples were removed from patients and stored at -80°C. The clinical data of the patients, including the age of onset, gender, tumor stage, tumor grade and other sample information, are shown in Table 1.
[0134] Table 1 Sample information
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[0137] During the biostatistical analysis, the 59 samples were divided into two parts. The first part was training modeling samples, including 19 bladder cancer samples and ...
Embodiment 2
[0139] Total RNA Extraction
[0140] About 50 mg of tissue fragments were excised from frozen tissues, and total RNA was extracted according to the instructions of miRNeasy mini kit (Qiagen, 217004).
[0141] 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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