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Aspergillus immunodominance antigen and application thereof

An immunodominant antigen, Aspergillus technology, applied in the field of immunodiagnosis, can solve problems such as little understanding

Inactive Publication Date: 2013-07-17
NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, little is known about these dominant antigenic components at home and abroad

Method used

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  • Aspergillus immunodominance antigen and application thereof
  • Aspergillus immunodominance antigen and application thereof
  • Aspergillus immunodominance antigen and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1 Patient Specimen Source, Diagnostic Criteria

[0052] 1. Sources of patient specimens 1,099 serum specimens from 781 patients were collected from the hematology, respiratory, ICU, oncology, nephrology, and rheumatology departments of 26 hospitals in Nanjing and surrounding areas. There were 484 males and 297 females, aged 18-99 years. The basic diseases of these patients are: malignant blood diseases such as acute and chronic leukemia, severe aplastic anemia, malignant lymphoma, etc. and 230 cases after hematopoietic stem cell transplantation, chronic lung diseases such as chronic obstructive pulmonary disease (COPD), chronic bronchitis, bronchiectasis 247 cases of pulmonary tuberculosis and severe pneumonia, 47 cases of autoimmune diseases such as systemic lupus erythematosus (SLE), dermatomyositis, rheumatoid arthritis, Sjogren's syndrome, etc., kidney diseases such as nephrotic syndrome, acute and chronic renal failure 36 cases of acute and chronic neph...

Embodiment 2

[0055] Example two Aspergillus fumigatus immunodominant antigen screening method

[0056] 1. Strains and culture conditions Aspergillus fumigatus A1 is a standard strain preserved by the Medical Fungi Center of China Committee for Culture Collection of Microorganisms, and the preservation number is CMCC(F)A1a. The fungus was cultured on Sandburg's agar medium at 28°C, the conidia were suspended in 0.9% NaCl (containing 0.1% Tween 80), and counted on a hemocytometer.

[0057] 2. The preparation of bacterial protein and secreted protein will be 2×10 7 Conidia of Aspergillus fumigatus were inoculated in YEPG medium respectively, and cultured with shaking at 150 rpm at 37°C for several days. The culture supernatant and hyphae were taken for the preparation of fungal antigens. Preparation of bacterial protein: filter the culture, collect mycelia, wash several times with PBS, grind thoroughly with liquid nitrogen, and extract protein with trichloroacetic acid (TCA) / acetone precipi...

Embodiment 3

[0094] Example Aspergillus trifumigatus Thioredoxin Reductase Gene Cloning

[0095]1. Strains, carriers and tool enzymes The standard strain of Aspergillus fumigatus A1 was obtained from the Medical Fungi Center of China Committee for Culture Collection of Microorganisms, and its preservation number is CMCC(F)A1a. ; pET28a (+) expression vector and PMD18-T cloning vector were purchased from Dalian Bao Biological Company, the host bacteria were E.coli JM109 and E.coli BL21(DE3); restriction enzymes Nde I and BamH I, T4 ligase , Plasmid Extraction Kit, DNA Fragment Purification Kit, and Gel Cut Recovery Kit were purchased from Dalian Bao Biology Company; Taq DNA polymerase, Total RNA Extraction Kit and RT-PCR Kit were purchased from Promega Company.

[0096] 2. Primer design and synthesis According to the Aspergillus fumigatus GliT coding sequence reported on Genbank, a pair of specific primers were designed using Primer 5 software, and a restriction enzyme site and several prot...

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Abstract

The invention discloses one or more immunodominant antigens of aspergillus selected from aspergillus thioredoxin reductase, fructose-bisphosphate aldolase, actin cytoskeleton protein VIP1, secreted dipeptidyl peptidase. The invention further discloses a use of the antigen for preparing diagnostic reagents for invasive aspergillosis, and diagnostic kits for invasive aspergillosis comprising the antigen.

Description

technical field [0001] The invention belongs to the field of immunodiagnosis, and relates to an aspergillus immunodominant antigen and its application. More specifically, the present invention relates to newly identified Aspergillus fumigatus antigens which strongly react with specific antibodies in the sera of patients with invasive aspergillosis. Through gene recombination technology, use prokaryotic cell expression system to obtain recombinant protein. The present invention also relates to the use of the immunodominant antigen in the serological diagnosis of invasive aspergillosis. Background technique [0002] Invasive aspergillosis (IA) is an infectious disease and is a serious fungal infection with high mortality. In recent years, the incidence of this infection has increased significantly, and it has become an important cause of death for patients with low immune function such as blood diseases, organ transplants, bone marrow transplants, AIDS, and autoimmune diseas...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02C12N9/88C12N9/48C07K14/38G01N33/569
CPCC07K14/38G01N33/56961G01N2333/38
Inventor 李芳秋史利宁
Owner NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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