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Application of latent membrane protein 2A of recombined EB virus for testing nasopharyngeal carcinoma

A latent membrane protein and Epstein-Barr virus technology, applied in biological testing, material inspection products, color/spectral characteristic measurement, etc., can solve problems such as cumbersome, poor repeatability, and unstable results

Inactive Publication Date: 2005-10-26
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are foreign reports using LMP2A expressed on the cell surface as an antigen to detect the LMP2A antibody in the serum of NPC patients by immunofluorescence method, but this method still has problems of tediousness, unstable results, and poor reproducibility

Method used

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  • Application of latent membrane protein 2A of recombined EB virus for testing nasopharyngeal carcinoma
  • Application of latent membrane protein 2A of recombined EB virus for testing nasopharyngeal carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1: Preparation of recombinant LMP2A

[0017] The preparation of recombinant LMP2A refers to the genetic engineering method in the article "Construction of Pichia pastoris expression vector containing EB virus LMP2A and detection of expressed LMP2A protein" (Journal of Nanjing Medical University, 2004, 24 (3)).

Embodiment 2

[0018] Example 2: Detection of anti-LMP2A antibody in the serum of patients with Epstein-Barr virus-related diseases

[0019] ELISA method detection:

[0020] Serum: 23 samples from EBV-negative healthy people (HN), 18 samples from EBV-positive healthy people (HI), 76 samples from EBV-related NPC patients (NPC), 8 samples from EBV-related Hodgkin's patients (HD), gastric cancer 15 patients (GC) and 24 other tumor patients not related to Epstein-Barr virus.

[0021] Steps:

[0022] (1) Antigen coating: the purified recombinant LMP2A antigen was diluted with 0.01M / L PBS to a concentration of 2.5ug / ml for coating. Add 100ul / ml antigen diluent to each well, overnight at 4°C.

[0023] (2) Plate washing: wash the microtiter plate 3 times with PBST (PBS containing 0.05% Tween), 3 minutes each time.

[0024] (3) Blocking: block with PBS containing 5% skimmed milk powder at 37° C. for 1 hour.

[0025] (4) Plate washing: Same as above.

[0026] (5) Primary antibody reaction: Epste...

Embodiment 3

[0035] Embodiment 3: Anti-VCA IgA and anti-LMP2A IgA antibody detection in NPC patient serum

[0036] 76 NPC patient sera were collected and tested for anti-VCA IgA and anti-LMP2A IgA antibodies respectively. The results are shown in Table 2.

[0037] VCA-IgA

(IF)

LMP2A-IgA

(ELISA)

+

-

Total (%)

+

-

39

3

14

20

53(70％)

23(30％)

Total (%)

42(55％)

34(45％)

76(100％)

[0038] Comparing the two results, P>0.05. More importantly, 3 samples were negative for anti-VCA IgA and positive for anti-LMP2A IgA. This indicates that there is a humoral immune response against these two virus antigens in patients with nasopharyngeal carcinoma, suggesting that the combined detection of antibodies against these two antigens can help improve the diagnosis of nasopharyngeal carcinoma patients.

[0039] The Western blots detection of the sera of the above t...

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Abstract

This invention relates to how to diagnose NPC. It shows the use of LMP2A-the membrane protein infected by recombined EBV in NPC diagnoses. It uses LMP2A as antigen, and detect serum sample by ELISA method. If the result is greater than the critical value, it is light demanding. This invention can be used to the early diagnoses of NPC, it is quick and convenient.

Description

technical field [0001] The invention relates to the detection of nasopharyngeal carcinoma, in particular to the application of recombinant Epstein-Barr virus latent membrane protein 2A (LMP2A) in the detection of nasopharyngeal carcinoma. Background technique [0002] Nasopharyngeal carcinoma (NPC) is a malignant tumor derived from epithelial cells. It exists widely all over the world. It is also a high-incidence tumor in southern my country such as Guangdong, Guangxi, Hong Kong and Fujian. There are more than 80,000 new cases. Radiotherapy and chemotherapy are the main means of treatment for NPC patients at present, and surgical treatment is usually not available, and radiotherapy has a poor therapeutic effect on advanced patients, with a survival rate of only 14%. Therefore, early diagnosis of patients is extremely important for improving the survival rate of NPC patients. important. [0003] Epstein-Barr virus (EBV) is closely related to the occurrence of NPC. Early diag...

Claims

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Application Information

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IPC IPC(8): G01N33/53
Inventor 姚堃陈云彭光勇孙华谢芳艺丁传林周峰吴文翰
Owner NANJING MEDICAL UNIV
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