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Microsatellite colorectal cancer instability amplification system and detection kit thereof

An unstable and amplification system technology, applied in the field of PCR amplification system, can solve limited problems and achieve the effects of convenient operation, cost saving, and large-scale promotion and application

Inactive Publication Date: 2014-02-05
上海赛安生物医药科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are many deficiencies in cost control and operability for multiple fluorescent labels, and the addition of sex-determining sites is also very limited in preventing experimental operators from confusing samples.

Method used

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  • Microsatellite colorectal cancer instability amplification system and detection kit thereof
  • Microsatellite colorectal cancer instability amplification system and detection kit thereof
  • Microsatellite colorectal cancer instability amplification system and detection kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The microsatellite instability detection kit for colorectal cancer of this embodiment consists of at least six test tubes containing corresponding reagents fixed in a paper packaging box, and the six test tubes are amplification buffer test tubes, Mg 2+ Test tubes, dNTP tubes, hot start enzyme tubes, primer mix tubes, and ultrapure water tubes. There is no special requirement for the order in which the above-mentioned test tubes are arranged in the packaging box, as long as they are clearly marked.

[0049] The development principle and method of use of the detection kit of the present embodiment are:

[0050] A. MSI gene locus primer design, synthesis and screening:

[0051] According to Bethesda standard microsatellite loci, including five genes BAT25, BAT26, D2S123, D5S346, and D17S250, the polymorphic microsatellite marker primers were designed.

[0052] Design primers using Oligo7.0 software for performance evaluation. Firstly, make the Tm value of each primer re...

Embodiment 2

[0097] The rest of the microsatellite instability detection kit for colorectal cancer in this embodiment is the same as that in Embodiment 1, except that the concentrations of the forward primer and the reverse primer in the universal-specific chimeric primer pair are different.

Embodiment 3

[0099] The rest of the microsatellite instability detection kit for colorectal cancer in this embodiment is the same as that in Embodiment 1, except that the concentrations of the forward primer and the reverse primer in the universal-specific chimeric primer pair are different.

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Abstract

The invention relates to a microsatellite colorectal cancer instability amplification system and a detection kit thereof. The amplification system disclosed by the invention comprises a general-specific chimeric primer pair and a mixture of general primers, wherein the general-specific chimeric primer pair respectively aims at microsatellite sites BAT25, BAT26, D2S123, D5S346 and D17S250; the general primer is FluD5-Up; respective positive primer 5' ends in the general-specific chimeric primer pair are provided with a sequence of the FluD5-Up; the FluD5-Up is a fluorescent-labeled primer. The amplification system and the detection kit thereof which are disclosed by the invention overcome the shortcoming that different primers require for multiple fluorescent labels; five microsatellite gene sites are detected in the same PCR (polymerase chain reaction) system, so that a result is intuitive and reliable; the time can be saved, and the efficiency is high; MSI (medium-scale integration) gene sites can be effectively detected and analyzed.

Description

technical field [0001] The invention relates to a PCR amplification system and detection products using the amplification system, belonging to the field of biotechnology. Background technique [0002] The incidence of colorectal cancer is gradually increasing both at home and abroad, and it is one of the common malignant tumors that seriously endanger human health. Studies have found that genome instability plays an important role in the pathogenesis of colorectal cancer. Genome instability includes chromosome instability (chromosomal instability, CI) and microsatellite instability (microsatellite instability, MSI). Previous studies on the incidence of colorectal cancer have shown that chromosomal instability (chromo-some instability) is the main cause of colorectal cancer, and chromosomal instability refers to an accelerated increase or decrease in the number of all chromosomes. In recent years, studies have found that microsatellite instability is another important mechan...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/156
Inventor 赵新泰王明
Owner 上海赛安生物医药科技股份有限公司
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