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Reagent kit used for detecting nasopharynx cancer and detection method of reagent kit

A kit and technology for nasopharyngeal carcinoma, applied in biological testing, biochemical equipment and methods, measuring devices, etc., can solve the problems of lack of sensitivity and specificity, and achieve the effects of low cost, easy preparation and easy preservation

Inactive Publication Date: 2016-09-28
张玲
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these indicators alone or in combination provide useful information for the diagnosis of nasopharyngeal carcinoma, they lack sufficient sensitivity and specificity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The acquisition of embodiment 1NASG protein

[0020] The NASG gene indicated by AF439448.1 was expressed by a conventional eukaryotic expression method in the art, and the corresponding target polypeptide protein was obtained.

Embodiment 2

[0021] Screening and preparation of embodiment 2 nucleic acid aptamers

[0022] Design a random nucleic acid library comprising approximately 20 nucleotides at both ends and 40 nucleotides in the middle as follows:

[0023] 5'-TAGCATGCAATGCCAGTATAG(N40)AACGTGCATGAACTATGAGT-3'; N40 represents 40 random nucleotides.

[0024] The single-stranded DNA library is amplified into double-stranded DNA, and the product is subjected to 2% agarose gel electrophoresis and then gel-cut to recover and purify; the recovered double-stranded DNA is used as a template to transcribe a single-stranded RNA random library in vitro, and the transcript is purified by PAGE . 75 μg RNA library was reverse-screened with nitrocellulose membrane to remove membrane-bound RNA molecules, and then incubated with 2ug NASG protein at 37°C for 30 min, the reaction solution was filtered through nitrocellulose membrane, and the filter membrane was washed; then the filter membrane was cut into pieces and placed in ...

Embodiment 3

[0055] Example 3 The performance measurement of protein binding suitable gametes

[0056] Take 2.0 μg of aptamers, digest them with calf intestinal alkaline phosphatase (CIP) at 37°C for 1 hour, purify and recover the dephosphorylated RNA; label [γ-32P]ATP on the dephosphorylated RNA by T4 polynucleotide kinase end of RNA molecule. 10nmol of radioactively labeled aptamers were incubated with different concentrations (1-200nM) of NASG at 37°C for 30min, the reaction solutions of each group were filtered through a nitrocellulose membrane, the filter membrane was washed, dried, and detected by a liquid scintillation counter. For the residual radioactivity, the same sample was measured twice in parallel. The dissociation constants of each aptamer and leptin were calculated. The result is as follows:

[0057] name

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Abstract

The invention discloses a reagent kit used for detecting nasopharynx cancer. An aptamer has good affinity to human NASG protein. NASG protein in the nasopharynx can be captured with the aptamer, and nasopharynx cancer can be detected according to the change of the content of NASG protein; the aptamer is prepared into the corresponding reagent kit to be applied to screening of nasopharynx cancer. The reagent kit has the advantages of being high in sensitivity, low in cost and easy to prepare and preserve.

Description

technical field [0001] The invention relates to a kit for detecting nasopharyngeal carcinoma and a detection method thereof. Background technique [0002] Nasopharyngeal carcinoma (NPC) is a common malignant tumor in southern my country, and Guangdong is a high-incidence area. Nasopharyngeal carcinoma refers to the cancer of nasopharyngeal mucosal epithelium, most of which are poorly differentiated squamous cell carcinoma, with high malignancy and hidden location, especially in the pharyngeal recess and the top of the nasopharynx. The early symptoms are not obvious, so it is difficult to early stage It was found that the rate of misdiagnosis and mistreatment is high, up to 12.2%. Therefore, in the diagnosed nasopharyngeal carcinoma, its 5-year survival rate hovers around 50% to 60% for a long time. Early diagnosis of nasopharyngeal carcinoma for early treatment and improving the survival rate of patients has always been one of the important subjects of NPC clinical research...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/574
CPCC12N15/115C12N2310/16G01N33/57407G01N33/57484G01N33/68G01N33/6812G01N33/6893G01N2800/14
Inventor 张玲
Owner 张玲