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

A kit and promoter 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 effects of low price, few influencing factors, and accurate detection.

Inactive Publication Date: 2016-05-18
苏州捷诺威生物科技有限公司
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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 weight of mutations other than C228T in the total mutations is about 1. %, and the mixed multiplex PCR cannot determine what kind of mutation the patient has, and sequencing is still needed to determine the specific mutation. If simple PCR is used, multiple PCR reactions need to be done, and the workload is heavy

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

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Embodiment 1

[0049] Embodiment 1: Detection of TERT gene promoter mutation

[0050] The kit for detecting TERT gene promoter mutations is a method for detecting TERT gene promoter mutations in non-disease diagnosis, comprising the following steps:

[0051] (1) Extract the urine of the sample to be tested, centrifuge to extract the urine cells, extract the whole DNA of the cells, and use the specific primers of SEQ ID NO.3-4 to carry out real-time PCR amplification of the template DNA;

[0052] Upstream primer: 5'-cccttcaccttccagctc-3' (SEQ ID NO.3);

[0053] Downstream primer: 5'-agcgctgcctgaaactgg-3' (SEQ ID NO.4);

[0054] (2) adding the fragment amplified in step (1) to the fluorescent probe of SEQ ID NO.1-2 for heating and melting;

[0055] The nucleotide sequence of the donor probe is as follows:

[0056] Donor probe: 5'-CGTCCCGACCCCTCCCGGGTC-Flc-3' (SEQ ID NO.1);

[0057] Receptor probe: 5'-Lc640-GCCCAGCCCCTTCCGGGCCCT-Phos-3' (SEQ ID NO.2);

[0058] (3) The mutation of the TERT ...

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Abstract

The invention belongs to a clinical medicine gene detection technique, and particularly relates to a kit for detecting TERT gene promoter mutation, and a detection method and application of the kit. The kit comprises a fluorescence labeled probe for the TERT gene. Only a pair of probes and a pair of primers are needed in the design, and the detected mutation is specific C228T mutation and accounts for 99% of the bladder cancer TERT mutation. The detection method has the advantages of simple reagents and low price; and the reaction system only contains the most basic primers and probes, so the influencing factors are fewer. The method is accurate for detection and simple to operate, and lays a solid foundation for quick detection of bladder cancer in clinical medicine.

Description

technical field [0001] The invention belongs to clinical medical gene detection technology, and in particular relates to a kit for detecting TERT gene promoter mutation, a detection method and 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-grade tumors, but i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q1/6858C12Q2600/156C12Q2531/113C12Q2527/107
Inventor 邓友平王红卫张绍波
Owner 苏州捷诺威生物科技有限公司
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