Polypeptide NPCT8 for NPC (Nasopharyngeal Carcinoma) screening, kit for NPC screening and application thereof

A kit and technology for nasopharyngeal carcinoma, applied in the field of nasopharyngeal carcinoma screening, can solve problems such as high missed diagnosis rate

Active Publication Date: 2020-02-14
同昕生物技术(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the existing Rta-IgG antibody detection kits have high sensitivity (average 87.8%) and specificity (average 92.8%), there is still a high rate of missed diagnosis

Method used

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  • Polypeptide NPCT8 for NPC (Nasopharyngeal Carcinoma) screening, kit for NPC screening and application thereof
  • Polypeptide NPCT8 for NPC (Nasopharyngeal Carcinoma) screening, kit for NPC screening and application thereof
  • Polypeptide NPCT8 for NPC (Nasopharyngeal Carcinoma) screening, kit for NPC screening and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Using a phage display peptide library to screen epitope peptides that bind to nasopharyngeal carcinoma serum antibodies

[0041] 1. Serum IgG antibody preparation in patients with nasopharyngeal carcinoma

[0042] Epstein-Barr virus Rta-IgG detection kit (Tongxin Biotechnology (Beijing) Co., Ltd., product number A-0496) was used to detect serum samples from 70 patients with nasopharyngeal carcinoma, and the detection steps were operated according to the kit instructions. 62 cases of Rta-IgG positive serum samples and 8 cases of Rta-IgG negative serum samples were obtained.

[0043]Eight cases of Rta-IgG negative serum samples were mixed, diluted with 2 times the volume of acetate buffer (0.06M, pH 4.8), added octanoic acid at 33 μl / ml serum, stirred and centrifuged to collect the supernatant. Ammonium sulfate was added to the supernatant at 0.277 g / mL, placed at 4°C overnight, centrifuged, and the supernatant was discarded. Add PBS buffer (0.01M, pH 7.2) to...

Embodiment 2

[0063] Example 2: Preparation of nasopharyngeal carcinoma detection kit

[0064] 1. Preparation of R185 protein

[0065] Rta protein (GenBank No. gi: 94734074) has a full length of 605aa, and its amino acid sequence is shown in SEQ ID NO: 11. The fragment of 171-356 amino acids was selected for preparation and named R185 protein, and its amino acid sequence is shown in Sequence 2. For the preparation method of R185 protein, refer to the examples 1 and 2 described in the instructions of the patent authorization text of the Chinese patent No. ZL200610113403.2 and the invention title "ELISA kit for screening, diagnosis and treatment effect prediction of nasopharyngeal carcinoma" ( [0098] on page 8 to [0202] on page 15), the main steps include the acquisition of the BRLF1 gene (GenBank No. gi: 94734074), the acquisition of the recombinant plasmid pGST-R185, the preparation of the GST-R185 protein and Purification, the purified GST-R185 protein is the R185 protein. This patent i...

Embodiment 3

[0080] Example 3: Sensitivity and specificity detection of nasopharyngeal carcinoma detection kit

[0081] Sera from 56 patients with nasopharyngeal carcinoma and 100 normal subjects were collected. Use the nasopharyngeal carcinoma detection kit of Example 2 (the coating ratio of NPCT8 and R185 protein on the enzyme plate is 1:1 mass ratio) and the control kit (EB virus Rta-IgG detection kit, Tongxin Biological , Product No. A-0496) for testing. The detection steps refer to the operation steps in the instructions of each kit respectively.

[0082] The test results are shown in Table 3 below. The specificity of the kit of the present invention is 95%, and the sensitivity is 95%. The control kit had a specificity of 93% and a sensitivity of 89%.

[0083] table 3

[0084]

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Abstract

The invention relates to NPC (Nasopharyngeal Carcinoma) screening, in particular to a polypeptide NPCT8 for NPC screening, a kit for NCP screening and application thereof. The amino acid sequence of the epitope peptide NPCT8 for NCP screening provided by the invention is shown as a sequence 1. The epitope peptide NPCT8 and an antigen protein shown as a sequence 2 are combined to be used for NPC screening; the sensitivity is 95 percent; and the specificity is 95 percent; and the early diagnosis rate of an NPC patient is further improved.

Description

technical field [0001] The present invention relates to the screening of nasopharyngeal carcinoma, in particular to a NPCT8 polypeptide specifically binding to the serum antibody of patients with nasopharyngeal carcinoma, an ELISA kit prepared by using the polypeptide, and the use of the polypeptide and the kit in the early screening of nasopharyngeal carcinoma Applications. Background technique [0002] Nasopharyngeal carcinoma (Nasopharyngeal carcinoma, NPC) is a malignant tumor that occurs in the nasopharynx. It originates in the epithelium of the nasopharyngeal mucosa. Characteristics of growth and early metastasis. Nasopharyngeal carcinoma accounts for 78.08% of head and neck malignancies. It accounts for 92.99% of upper respiratory tract cancers. Nasopharyngeal carcinoma is common in young and middle-aged men, and the age of incidence is between 35 and 50, accounting for 60% of the incidence. On September 9, 2015, the National Health and Family Planning Commission ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08G01N33/574G01N33/68
CPCC07K7/08G01N33/57407G01N33/6854
Inventor 张建珍李全焦守恕吴凡
Owner 同昕生物技术(北京)有限公司
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