Compound amplification system and kit for detecting chromosome deficiency

A compound amplification system and chromosome deletion technology, which is applied in the field of PCR amplification system, can solve the problems of inconvenient and accurate determination of test results, complicated operation, and many doping factors in result analysis, so as to facilitate standardized operation and operation Convenience and conditional mode effects

Active Publication Date: 2014-05-21
国九堂山东阿胶有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the complicated operation of restriction enzyme digestion method and silver staining gel electrophoresis method, and there are many contamination factors in the result analysis, the test results cannot be judged conveniently and accurately.

Method used

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  • Compound amplification system and kit for detecting chromosome deficiency
  • Compound amplification system and kit for detecting chromosome deficiency
  • Compound amplification system and kit for detecting chromosome deficiency

Examples

Experimental program
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Effect test

Embodiment 1

[0047] The detection kit for detecting chromosomal deletion in this embodiment is composed 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.

[0048] The development principle and method of use of the detection kit of the present embodiment are:

[0049] A. Design, synthesis and screening of primers for glioma 1p19q LOH gene locus:

[0050] Primers were designed according to the six genes (D1S468, D1S436, D1S489, D19S112, D19S217, D19S902) of glioma 1p19q recommended by the International Cancer Society. Collect venous whole blood (anticoagulated whole blood) and tissue samples (fresh tumor tissue or paraffin section) from ...

Embodiment 2

[0094] The remaining parts of the compound detection kit for detecting chromosomal deletions in this embodiment are the same as in Example 1, except that the concentrations of the forward primer and the reverse primer in the universal-specific chimeric primer pair are different.

Embodiment 3

[0096] The remaining parts of the compound detection kit for detecting chromosomal deletions in this embodiment are the same as in Example 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 compound amplification system and a kit for detecting chromosome deficiency. The compound amplification system comprises a mixture of a universal-specific chimeric primer pair and a universal primer, wherein the universal-specific chimeric primer pair respectively aims at genetic locus of D1S468, D1S436, D1S489, D19S112, D19S217 and D19S902. The universal primer is FluD5-Up. A sequencer of FluD5-Up is arranged at the 5' end of each forward primer in the universal-specific chimeric primer pair, and the FluD5-Up is a fluorescence labeling primer. The compound amplification system and the detection kit provided by the invention overcome the deficiency that various fluorescence labels are required for different primers, and 6 1p19q genetic locus can be detected once by using a single fluorescence label in a PCR (Polymerase Chain Reaction) system, so that the system is intuitive and reliable in result and time-saving and efficient, and can effectively detect and analyze loss of heterozigosity of the chromosome 1p19q.

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] Tumors derived from the neuroepithelium are collectively called gliomas (gliomas), which account for 40-50% of brain tumors and are the most common intracranial malignant tumors, with an annual incidence of about 3-8 cases per 100,000 population. The World Health Organization (WHO) divides glioma into 4 grades, ranging from low to high malignancy, WHO grade I is benign, WHO grade II is low grade malignancy, and WHO grade III and WHO IV grades are highly malignant. In recent years, molecular biology research on the occurrence and development of glioma has been deepened. The neuropathological evaluation of glioma is no longer limited to providing clinical information about the histological type and malignant grade of the tumor. It may require more and more More convenient detec...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2537/143C12Q2527/143C12Q2565/125
Inventor 赵新泰王明
Owner 国九堂山东阿胶有限公司
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