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Mumps virus HN antigen and application of antigen in detection of mumps-resisting virus antibody

A mumps virus, antibody technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as poor sensitivity

Active Publication Date: 2017-01-04
北京市华信行生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the ELISA kits currently available on the market are coated with whole virus antigens, so the sensitivity is poor

Method used

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  • Mumps virus HN antigen and application of antigen in detection of mumps-resisting virus antibody
  • Mumps virus HN antigen and application of antigen in detection of mumps-resisting virus antibody
  • Mumps virus HN antigen and application of antigen in detection of mumps-resisting virus antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Selection of Candidate Antigen Neutralizing Epitopes

[0024] The present inventor carried out antigenicity prediction to mumps virus HN sequence by bioinformatics software, the result is as follows figure 1 As shown, there are 4 major antigenic segments whose sequences are shown in SEQ ID NO.1, 2, 3 and 4, respectively.

Embodiment 2

[0025] Example 2 Cloning, expression and activity identification of three mumps virus HN antigens

[0026] 1. Design and synthesis of HN antigen gene

[0027] In order to facilitate the efficient expression of the mumps virus HN gene in E.coli, based on the amino acid sequences of the four mumps virus HN antigens determined in Example 1, the biased codons of Escherichia coli were selected to design the HN gene, and commissioned Shanghai Yingwei Jieji Biotechnology Service Co., Ltd. synthesized the gene of the above sequence.

[0028] 2. Construction and transformation of HN antigen expression plasmid

[0029] In order to facilitate gene cloning into the expression vector, two restriction sites Xho I and Xba I were introduced at the two ends of the connecting arms of the two fragments, so as to be suitable for the expression vector pBVIL1 (see Chinese patent ZL00100695.9: expression vector pBVIL1 and its construction method and application), the two ends of the tether of the...

Embodiment 3

[0045] Example 3 Detection of mumps virus antibody

[0046]HN antigen 2 (SEQ ID NO.2) and antigen 3 (SEQ ID NO.3) were used to coat the enzyme-linked plate, and the neutralizing antibody of the mumps vaccine was detected by the indirect ELISA method using the goat anti-human IgG antibody, and the measurement results and The analysis results are shown in Table 3: Among the 90 cases of mumps before vaccination, 3 HN-IgG test results were positive for antigen 2, and 87 cases were negative, with a positive rate of 3.3%; the positive rate of antigen 3 was 8.9%. Among the 40 vaccination groups, 33 cases of HN-IgG were positive for antigen 2 and 7 cases were negative, the positive rate was 82.5%; the positive rate for antigen 3 was 95%. The positive rate of the vaccine group was significantly higher than that of the pre-epidemic group (P<0.01). Because the positive detection of antigen 2 and 3 is complementary, antigen 2 and 3 have high specificity and sensitivity as antibody detect...

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Abstract

The invention discloses a mumps virus HN antigen capable of detecting a mumps-resisting virus antibody, especially a mumps-resisting virus neutralized antibody and further discloses application of the antigen in preparation of detection agent for detecting the mumps-resisting virus neutralized antibody and a corresponding detection kit.

Description

technical field [0001] The invention relates to the field of immunodetection of antibodies, in particular to the field of immunodetection of anti-mumps virus antibodies, especially neutralizing antibodies. Background technique [0002] Mumps is an acute infectious disease caused by Mumps virus (MV). The transmission of mumps virus is usually carried by air droplets, which also leads to the high transmission rate of the disease. Under normal circumstances, the possibility of obvious clinical symptoms after infection with mumps virus is about 60%, including fever, loss of appetite, headache, general malaise and other symptoms, and the subsequent bilateral or unilateral parotid gland enlargement has diagnostic significance . Mumps, as an acute respiratory infectious disease characterized by parotid gland enlargement, mostly occurs in children and young adults, and its main hazards are complications such as encephalitis, meningitis, and orchitis. Among the infected people, 5%...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983G01N2333/12
Inventor 张贺秋孙妮娜陈庆全
Owner 北京市华信行生物科技有限公司
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