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Kit for colorectal cancer detection and its application

A colorectal cancer and kit technology, applied in the fields of biotechnology and medicine, can solve the problem that the sensitivity cannot meet the clinical needs, and achieve the effect of improving the 5-year survival rate, high sensitivity, and reducing false negatives

Active Publication Date: 2020-01-10
GENETALKS BIO TECH CHANGSHA CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] At present, the only product in my country for the detection of methylation of colorectal cancer-related genes is SEPT9 methylation detection based on plasma samples. Although its specificity is high, its sensitivity is far from meeting clinical needs.

Method used

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  • Kit for colorectal cancer detection and its application
  • Kit for colorectal cancer detection and its application
  • Kit for colorectal cancer detection and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] A kit for detecting colorectal cancer of the present invention, comprising a primer composition, a probe composition and a blocking sequence composition, 2x 360 Master Mix, tetramethylammonium chloride (TMAC), glycerol and deionized water.

[0084] The components of the primer composition are listed in Table 1.

[0085] Table 1: Primer compositions of Example 1

[0086] BS-S9-F 5'-TTTGTATTGTAGGAGCGCGGGC-3' (SEQ ID NO.1) BS-S9-R 5'-CAACGACGAAAAAACGCCCCCG-3' (SEQ ID NO.2) BS-N4-F 5'-GTTTCGCGTCGCGGTTTTCGTTC-3' (SEQ ID NO.3) BS-N4-R 5'-CGTAACTTCCGCCTTCTACGCG-3' (SEQ ID NO.4) BS-SDC2-F 5'-AGAAATTAATAAGTGAGAGGGCGTCGC-3' (SEQ ID NO.5) BS-SDC2-R 5'-AACGCTCGCTTCCTCCTCCTACG-3' (SEQ ID NO.6) BS-ACT-F 5'-GAAAGGGTGTAGTTTTGGGAGGTTAG-3' (SEQ ID NO.7) BS-ACT-R 5'-CCCAAAACCAACCACAAAAAAAAT-3' (SEQ ID NO.8)

[0087] In Table 1, the primer pair BS-S9-F and BS-S9-R are used to amplify the SEPT9 gene; the primer pair BS-N4-F an...

Embodiment 2

[0098] An application of the kit of Example 1 in the detection of colorectal cancer, the application method comprising the following steps:

[0099] Sampling: Collect 5-8g of stool samples and put them into 15mL of stool sample protection solution. The components of the stool sample protection solution are listed in Table 4.

[0100] Table 4: Component list of stool sample protection solution

[0101] components concentration tris-HCl 0.5M Guanidine isothiocyanate 1.5M EDTA 10mM Sodium dodecyl sulfate SDS 0.2% (m / v) Triton X-100 1% (v / v) Tween 20 1% (v / v) glycerin 5% (v / v)

[0102] (2) Extraction: Extraction was carried out using Jinmag Fecal Genome Extraction Kit (magnetic bead method) from Renhe Future Biotechnology (Changsha) Co., Ltd. The specific steps are as follows:

[0103] 2.1. Fully oscillate and mix the stool sample (about 20mL) to a homogenized state, centrifuge at 3000g for 15min, transfer 10mL of the...

Embodiment 3

[0149] Investigate the sensitivity, specificity and accuracy of the method of embodiment 2. Based on the prediction model in Example 2, predictions were made on 90 newly-inspected samples, and compared with the actual values, a sample confusion matrix was obtained. Table 7 below is the sample matrix.

[0150] Table 7: Sample matrix table

[0151]

[0152]

[0153] Calculate the sensitivity, specificity and accuracy of the prediction model of the present application according to the predicted value and actual value in Table 7.

[0154] Wherein, sensitivity=true positive / (true positive+false negative)×100%;

[0155] Specificity = true negative / (true negative + false positive) × 100%;

[0156] Accuracy rate = (true positive + true negative) / total number of detections × 100%.

[0157] It can be seen from the results in Table 7 that the prediction model of Example 2 has a sensitivity of 0.8857; a specificity of 0.8909; an accuracy of 0.8889; and a Kappa of 0.7686.

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Abstract

The invention discloses a kit for colorectal cancer detection and an application thereof. The kit comprises a primer composition, and a probe composition. The primer composition comprises BS-S9-F, BS-S9-R, BS-N4-F, BS-N4-R, BS-SDC2-F, BS-SDC2-R, BS-ACT-F and BS-ACT-R. The probe composition comprises BS-S9-P, BS-N4-P, BS-SDC2-P and BS-ACT-P. The kit of the invention can be applied to the detectionof colorectal cancer, and has the advantages of high sensitivity and high specificity.

Description

technical field [0001] The invention relates to the fields of biotechnology and medicine, in particular to a kit for detecting colorectal cancer and its application. Background technique [0002] Cancer has become the biggest killer that threatens human health. The 2018 China cancer statistics show that in 2014, there were 3.804 million new cancer cases nationwide, an increase of 120,000 cases or 3.2% compared with 2013. In addition, Wanqing Chen predicted in his published "Cancer Statistics in China, 2015" that in 2015, the number of new cancer patients in my country will reach 4.292 million, and the number of new cancer deaths will reach 2.814 million. [0003] Colorectal cancer is a common malignant tumor of the digestive tract. There are no obvious symptoms in the early stage, and the patients have no obvious abnormal changes. As a result, many patients miss the best time for diagnosis and treatment. Once diagnosed, the cancer is already in the middle and late stages. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6886C12Q2600/154C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 宋卓王永利孙海鹏高堂杰周巧
Owner GENETALKS BIO TECH CHANGSHA CO LTD
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