Kit for detecting mutation of TERT gene promoter and detection method and application thereof

A promoter and kit technology, applied in the field of clinical medical gene detection, can solve the problems of complex reaction system of the kit, many influencing factors, heavy workload, etc., and achieve the effect of improving detection accuracy, reducing influencing factors, and simple reagents

Inactive Publication Date: 2016-11-23
上海汇真生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the reaction system of the kit is complex and there are many influencing factors. Each factor can affect the reaction result and interpretation, which is difficult to interpret; it seems that almost all mutations can be detected, but the mixed multiplex PCR cannot determine the mutation of the patient, and sequencing is still needed To determine the specific mutation, if you use simple PCR, you need to do multiple PCR reactions, and the workload is heavy

Method used

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  • Kit for detecting mutation of TERT gene promoter and detection method and application thereof
  • Kit for detecting mutation of TERT gene promoter and detection method and application thereof
  • Kit for detecting mutation of TERT gene promoter and detection method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] A kit for detecting mutations in the promoter of the TERT gene, comprising:

[0071] (1) A fluorescently labeled probe that specifically recognizes the mutation at the C228T site of the TERT gene promoter, the nucleotide sequences of the donor probe and the acceptor probe are as follows:

[0072] Donor probe for C228T mutation: 5'-CGTCCCGACCCCTCCCGGGTC-Flc-3'

[0073] Receptor probe for C228T mutation: 5'-Lc640-GCCCAGCCCCTTCCGGGCCCT-Phos-3'

[0074] (2) Specific primers for detecting the C228T site mutation of the TERT gene promoter, the nucleotide sequence of which is as follows:

[0075] Upstream primer: 5'-CCCTTCACCTTCCAGCTC-3'

[0076] Downstream primer: 5'-AGCGCTGCCTGAAACTGG-3'

[0077] (3) Master Mix, which includes: 1×PCR buffer, 0.2μM dNTPs, 1.5U / mg one Taq DNA polymerase and 10ng DNA template;

[0078] (4) An instruction manual.

Embodiment 2

[0080] A kit for detecting mutations in the promoter of the TERT gene, comprising:

[0081] (1) A fluorescently labeled probe that specifically recognizes the mutation at the C250T site of the TERT gene promoter, the nucleotide sequences of the donor probe and the acceptor probe are as follows:

[0082] Donor probe for C250T mutation: 5'-CGTCCCGACCCCTTCCGGGTC-Flc-3'

[0083] Receptor probe for C250T mutation: 5'-Lc640-GCCCAGCCCCCTCCGGGCCCT-Phos-3'

[0084] (2) Specific primers for detecting mutations at the C250T site of the TERT gene promoter, the nucleotide sequence of which is as follows:

[0085] Upstream primer: 5'-CCCTTCACCTTCCAGCTC-3'

[0086] Downstream primer: 5'-AGCGCTGCCTGAAACTGG-3'

[0087] (3) Master Mix, which includes: 1×PCR buffer, 0.2μM dNTPs, 1.5U / mg one Taq DNA polymerase and 10ng DNA template;

[0088] (4) An instruction manual.

Embodiment 3

[0090] A kit for detecting mutations in the promoter of the TERT gene, comprising:

[0091] (1) A fluorescently labeled probe that specifically recognizes the mutation at the C228T site of the TERT gene promoter, the nucleotide sequences of the donor probe and the acceptor probe are as follows:

[0092] Donor probe for C228T mutation: 5'-CGTCCCGACCCCTCCCGGGTC-Flc-3'

[0093] Receptor probe for C228T mutation: 5'-Lc640-GCCCAGCCCCTTCCGGGCCCT-Phos-3'

[0094] (2) A fluorescently labeled probe that specifically recognizes the mutation at the C250T site of the TERT gene promoter, the nucleotide sequences of the donor probe and the acceptor probe are as follows:

[0095]Donor probe for C250T mutation: 5'-CGTCCCGACCCCTTCCGGGTC-Flc-3'

[0096] Receptor probe for C250T mutation: 5'-Lc640-GCCCAGCCCCCTCCGGGCCCT-Phos-3'

[0097] (3) Specific primers for detecting mutations at the C250T site of the TERT gene promoter, the nucleotide sequence of which is as follows:

[0098] Upstream pr...

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Abstract

The invention belongs to the technology of clinical medicine gene detection and particularly relates to a kit for detecting mutation of a TERT gene promoter and a detection method and application thereof. The kit comprises a fluorescence labeling probe specifically recognizing mutation of a lotus C228T and mutation of a lotus C250T or other combinations of the TERT gene promoter, a pair of TERT specific primers and needed Master mix of all reagents. The specificity mutation C228T and the specificity mutation C250T of the TERT gene promoter are provided, and the sum of the specificity mutation C228T and the specificity mutation C250T accounts for about 100% of bladder cancer TERT mutation; a detection method is standardized, the reagents are subjected to optimization verification, the price is low, all ingredients in a reaction system are premixed, the operation process is simplified to the maximum extent, and operation errors are avoided. When the kit is used for detecting mutation of the TERT gene promoter, the number of influence factors is small, detection is precise, operation is easy, detection results are understood easily, and a reliable method for quickly detecting bladder cancer through urine is provided for clinical medicine.

Description

technical field [0001] The invention belongs to the technical field of clinical medical gene detection, and in particular relates to a kit for detecting TERT gene promoter mutation, a detection method and an application thereof. Background technique [0002] Urothelial carcinoma of the bladder is the most common malignancy of the urinary tract. The incidence rate of cancer in the 32 tumor registries across the country is 6.69 / 100,000, accounting for 2.52% of all new cases of malignant tumors. According to gender statistics, the incidence rates of bladder cancer in males and females are 10.10 / 100,000 and 3.20 / 100,000, respectively, and males are 3.16 times higher than females. The mortality rate of bladder cancer was 2.53 / 100,000 in 32 tumor registries nationwide, accounting for 1.47% of all malignant tumor deaths. Most histologically, this carcinoma is a well-differentiated papillary lesion. Urine cytology has high sensitivity and specificity in the diagnosis of high-grad...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 邓友平王红卫张绍波
Owner 上海汇真生物科技有限公司
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