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Marker related to abnormal pancreaticobiliary confluence and application thereof

A technology of pancreaticobiliary and markers, applied in the field of genetic engineering, can solve the unrecorded problems related to the occurrence of lncRNA and PBM, and achieve the effect of low cost, high throughput and strong specificity

Pending Publication Date: 2021-10-22
SOOCHOW UNIV AFFILIATED CHILDRENS HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, there is no record in the prior art that lncRNA is associated with the occurrence of PBM

Method used

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  • Marker related to abnormal pancreaticobiliary confluence and application thereof
  • Marker related to abnormal pancreaticobiliary confluence and application thereof
  • Marker related to abnormal pancreaticobiliary confluence and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] In this embodiment, the screening of gene markers related to pancreaticobiliary junction abnormality includes the following steps:

[0057] (1) Sample collection: Following the principle of informed consent and with the consent of the ethics committee, the common bile duct tissue and normal common bile duct tissue of patients with abnormal pancreaticobiliary junction were collected and stored at -80°C;

[0058] (2) RNA sample preparation: use TRIZOL reagent (Invitrogen, Carls-bad, CA, USA) to extract the total RNA in the tissue collected in step (1) according to the instructions, and use an ultra-micro spectrophotometer (NanoDrop ND-2000, ThermoScientific) to conduct Quantification of total RNA;

[0059] (3) Reverse transcription and labeling: Use the random primer method (Arraystar Super RNALabeling Kit; Arraystar) to reverse transcribe the RNA in step (2) to obtain cDNA, then use the cDNA to synthesize cRNA, and add Cyanine-3- CTP marker;

[0060] (4) Hybridization:...

Embodiment 2

[0066] The present embodiment extracts RNA in the sample, comprises the following steps:

[0067] (1) Samples: Abnormal pancreaticobiliary junction tissues and normal intestinal tissues from 15 patients with pancreaticobiliary junction abnormalities were collected following the principle of informed consent and with the consent of the ethics committee;

[0068] (2) RNA sample extraction:

[0069] 1) Reagents include TRIZOL reagent (Invitrogen life technologies), chloroform (Shanghai Chemical Reagent Co., Ltd.), isopropanol (Shanghai Chemical Reagent Co., Ltd.), 100% ethanol (Shanghai Chemical Reagent Co., Ltd.), 75% ethanol (treated with DEPC water), RNase-free water and RNase-free glycogen (Invitrogen lifetechnologies);

[0070] 2) Take 70 mg of tissue sample, add 1 mL of TRIZOL reagent, and homogenize with an electric homogenizer;

[0071] 3) Incubate the homogenized sample at 20°C for 5 minutes, add 0.2mL chloroform to each 1mL TRIZOL reagent homogenized sample, close the...

Embodiment 3

[0076] In this example, the RNA prepared in Example 2 was used for cDNA synthesis.

[0077] Reagents: RNase inhibitor (Epicentre), SuperScript TM III Reverse Transcriptase (Invitrogen), 5×RT buffer (Invitrogen), 2.5mM dNTP mixture (dATP, dGTP, dCTP and dTTP each 2.5mM) (HyTest Ltd) and Primer (Yingjun Biotechnology Co., Ltd.).

[0078] Instruments: clean bench (Shanghai Boxun Industrial Co., Ltd. Medical Equipment Factory), DK-8D electric heating constant temperature water tank (Shanghai Senxin Experimental Instrument Co., Ltd.), and GeneAmp PCR System 9700 (Applied Biosystems).

[0079] The operation process includes the following steps:

[0080] (1) Preparation of annealing mixture:

[0081]

[0082] (2) Place the mixture in a water bath at 65°C for 5 minutes, and place it on ice for 2 minutes. After centrifugation, add the RT reaction solution to the centrifuge tube in sequence:

[0083]

[0084] (3) After mixing, keep the temperature at 37°C for 1min, pipette gen...

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Abstract

The invention relates to a marker related to abnormal pancreaticobiliary confluence and application of the marker. The marker related to the occurrence of the pancreaticobiliary confluence abnormality comprises a long-chain non-coding RNAXR_946886. According to the kit, a molecular diagnosis technology is utilized, high-specificity XR_946886 is adopted as the marker for the pancreaticobiliary confluence abnormity for the first time, the marker not only can make diagnosis of the pancreaticobiliary confluence abnormity meet the requirements for high specificity and high sensitivity, but also has the multiple comprehensive advantages of being easy and rapid to operate, free of intervention, high in flux, low in cost and the like.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to a marker related to abnormal pancreaticobiliary juncture and its application, in particular to LncRNA (XR_946886) as a marker for abnormal pancreaticobiliary juncture. Background technique [0002] The junction of the common bile duct, the main pancreatic duct and the duodenal papilla is a subtle structure in the human body. The connection mode of the pancreaticobiliary duct also varies greatly among different individuals. About 85% of the common bile duct and the main pancreas The tube joins at an acute angle in the inner section of the duodenal wall to form a "common channel", which maintains the normal secretion pressure of the biliary tract and pancreatic juice, and prevents the pancreatic juice and duodenal contents from flowing back into the bile duct. If the bile duct and pancreatic duct are in the duodenum The intestinal walls meet at a right angle, making the "...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/113
CPCC12Q1/6883C12Q2600/158
Inventor 郭万亮汪健黄顺根赵廉
Owner SOOCHOW UNIV AFFILIATED CHILDRENS HOSPITAL
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