Molecular marker for early pancreatic neoplasm detection as well as detection method and application thereof
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A molecular marker, pancreatic tumor technology, applied in the field of biotechnology and clinical molecular diagnostic drug development, can solve the problems of difficult early diagnosis of pancreatic cancer, low sensitivity and specificity of biomarkers, etc.
Active Publication Date: 2020-08-25
KANTE DALIAN BIOTECHNOLOGIES INC
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[0011] Aiming at the current difficulty in early diagnosis of pancreatic cancer and the low sensitivity and specificity of existing biomarkers, the present invention provides a new method for screening and detecting pancreatic cancer with differential expression of miRNAs in serum and early pancreatic tumor molecular New uses for markers
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Embodiment 1
[0075] Early screening of miRNAs and construction of precise drug detection system (SOP)
[0076] S1. To screen pancreatic cancer miRNAs, design specific probes and primers for target miRNAs according to the principle of TaqMan probe technology; react target miRNAs in U6 in the same system; the designed primers and probes are as follows:
[0077] Table 1. Design of primers and probe sequences for target miRNAs
[0081] (1) Determine the extraction reagent. Screened three commonly used commercial RNA isolation kits, namely TRIzol (Ambion), TRIzol LS Reagent (Invitrogen) and miRNeasy Serum / Plasma kit (Qiagen), compared the quality of the extracted miRNA, and evaluated the cost and ease of operation degree and other factors, select the extraction reagent with the best comprehensive performance. The three extraction reagents were used to extract the same test sample (taking Sw1990 pancreatic cancer cel...
Embodiment 2
[0096] Utilizing the multiple fluorescent probe detection technology for miRNA in serum established by the present invention, miR-30c, miR-24, miR-23a, miR-132, miR-21, let The expression of -7i, miR-1207, miR-130, miR-200c, miR-154, and miR-57 were divided into 6 combinations, namely,
[0103] Specifically: serum samples from patients from Shanghai Renji Hospital, Dalian Medical University, Peking Union Medical College Hospital and Nanjing Medical University (including early pancreatic ductal adenocarcinoma, pancreatitis, pancreatic ductal papillary mucinous tumor) and normal samples from Hunan Xiangya Hospital Human serum samples are operat...
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Abstract
The invention discloses a molecular marker for early pancreatic neoplasm detection as well as a detection method and application thereof and belongs to the field of biotechnology and clinical molecular diagnosis drug development. A pancreatic cancer marker miRNAs includes miR-30c, miR-24, miR-23a and miR-132. Combination, a method and a kit provided by the invention can be used for evaluation on aspects of screening and identifying diagnostics of early pancreatic cancer, monitoring of disease complications and recurrence, curative efficacy, efficacy, accurate medication guidance and the like and have the advantages of wide detection lineage range, high sensitivity, good specificity, low detection cost, availibility in material, easies in storage of samples and the like. The method can be widely used for related work such as early survey and prognosis of pancreatic cancer, improves individual differences difficult to overcome as well as low specificity and low sensitivity brought by theinstability of a single marker or a biomarker which is clinically widely applied self at present, significantly increases the clinical detection rate of the early pancreatic cancer, lowers the misdiagnosis rate and the missed diagnosis rate of the pancreatic cancer and becomes effective means of screening and diagnosis of the early pancreatic cancer.
Description
technical field [0001] The invention belongs to the field of biotechnology and clinical molecular diagnosis drug development, and particularly relates to the ability to distinguish early pancreatic cancer, pancreatitis and intraductal papillary mucinous tumor lesions of the pancreas and normal human blood detection markers, PCR (polymerase chain reaction) or any of these The marker is a method and a kit for detection means. Background technique [0002] Early pancreatic cancer usually refers to pancreatic cancer with a tumor diameter ≤ 2.0 cm, no lymph node metastasis, no pancreatic capsule and peripancreatic invasion, no invasion of blood vessels and adjacent organs, and the stage is T1aN0M0. However, some scholars believe that most pancreatic cancers of 1.0 cm to 2.0 cm have metastasized to lymph nodes, and advocated that the tumor diameter ≤ 1.0 cm is the standard for early pancreatic cancer. Unless the lesion happens to be located in the duodenal papilla, symptoms of bi...
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