Non-invasive detection method and kit for early screening bladder cancer by multi-gene combination

A bladder cancer and kit technology, applied in the field of molecular biology, can solve the problems of large number of target genes, many primers, discomfort, etc., and achieve the effect of improving sensitivity and specificity, high accuracy, and improving sensitivity and specificity

Pending Publication Date: 2018-09-14
ANHUI DAJIAN MEDICAL TECH CO LTD
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Problems solved by technology

The combined detection of several genes is the trend of methylation detection in bladder cancer, but whether these reported methylation markers can be used to improve diagnostic sensitivity and specificity is still a problem (Wang Yongqiang, urine EOMES, PCDH17 and Significance of POU4F2 gene methylation analysis for early diagnosis of bladder urothelial carcinoma), Catto et al. stud...

Method used

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  • Non-invasive detection method and kit for early screening bladder cancer by multi-gene combination
  • Non-invasive detection method and kit for early screening bladder cancer by multi-gene combination
  • Non-invasive detection method and kit for early screening bladder cancer by multi-gene combination

Examples

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Embodiment 1

[0047] Example 1 Design of primers and probes and their kits for detecting the methylation level of multiple genes in urinary sediment cells related to bladder cancer.

[0048] The present invention simultaneously designed and verified multiple primer and probe combination sequences for the methylated nucleic acid sequences of three target gene fragments related to bladder cancer, APC, NID2, and p16, and optimized the following three sets of primer probe sequences:

[0049] (1) Detect the nucleic acid sequence of APC gene fragment methylation (FAM fluorescent channel)

[0050] The forward primer sequence is SEQ ID NO: 1,

[0051] The reverse primer sequence is SEQ ID NO: 2,

[0052] The fluorescent probe sequence is SEQ ID NO: 3;

[0053] Fluorescent probe SEQ ID NO:3; the 5' end of the fluorescent probe SEQ ID NO:3 is labeled with FAM (fluorescence reporter group), and the 3' end is labeled with MGB (fluorescent quencher group).

[0054] (2) Detect the nucleic acid sequenc...

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Abstract

The invention discloses a non-invasive detection method and a kit for early screening a bladder cancer by multi-gene combination. The combined genes include APC, NID2 and p16, and specific methylationprimers are designed to detect three nucleotide sequences of target gene fragment methylation of the APC, the NID2 and the p16, so that early screening and auxiliary diagnosis of the bladder cancer are achieved. Methylation levels of three genetic markers of the APC, the NID2 and the p16 in urinary sediments are analyzed by a three-channel fluorescent quantitative PCR (polymerase chain reaction)method, and a positive or negative result of a sample is judged according to a reported CT (computed tomography) value. The non-invasive detection method can detect 90% of early bladder cancers, 64% of precancerous adenomas (larger than or equal to 1 centimeter) in preclinical study, specificity reaches 100%, invasiveness is completely omitted, the method only needs to collect urine of 50mL, and acceptability of throngs wearing no symptoms is high.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to a non-invasive detection method for multi-gene joint screening of bladder cancer and a kit thereof, which are suitable for detecting whether there is bladder cancer mutation DNA in urine sedimentation cells. Background technique [0002] Bladder cancer (BC) is one of the most common malignant tumors of the urinary system, accounting for the first place in urinary system tumors, more than 90% of which are transitional cell carcinomas (BTCC), of which 80% are invasive superficial carcinomas, which are cured for the first time The subsequent recurrence rate is 70% to 80%, and its incidence and recurrence rate are increasing year by year. The incidence rate of bladder cancer varies by as much as 10 times among countries in the world. Western Europe and North America are the highest, while Eastern Europe and some Asian countries are relatively low. In my country, the incidence of male b...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/118C12Q2600/156
Inventor 邵琦
Owner ANHUI DAJIAN MEDICAL TECH CO LTD
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