Immune Netw.  2008 Sep;8(3):82-89. 10.4110/in.2008.8.3.82.

Detection of Serum IgE Specific to Mite Allergens by Immuno-PCR

Affiliations
  • 1Division of Molecular and Medical Biotechnology, College of Bioscience and Biotechnology, Kangwon National University, Chuncheon, Korea. chash@kangwon.ac.kr
  • 2Department of Paediatrics, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
  • 3Member of Immunology Programme, National University of Singapore, Singapore.
  • 4Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research, Singapore.
  • 5Institute of Bioscience and Biotechnology, Kangwon National University, Chuncheon, Korea.
  • 6IG Therapy Co., Rm 112, Nong 3rd Building, Kangwon National University, Chuncheon, Korea.

Abstract

BACKGROUND: Although a skin test is the primary option for detecting allergen-specific IgE in clinics, the serum IgE immunoassay is also important because it allows for the diagnosis of allergy without any accompanying adverse effect on the patient. However, the low detection limit of IgE levels by immunoassay may restrict the use of the method in some occasions, and improving its sensitivity would thus have a significant implication in allergy-immunology clinics.
METHODS
In this study, we attempted to detect specific serum IgE by using immuno-polymerase chain reaction (IPCR) which combines the antigen-antibody specificity of enzyme-linked immunosorbent assays (ELISAs) with the amplification power of PCR.
RESULTS
Our results demonstrated that Blo t5-specific serum IgE can be detected by IPCR with a 100-fold higher sensitivity than ELISA, and cross-reactivity of serum IgE to other mite allergens is able to be analyzed by using only 0.3 microliter of serum sample. Use of real-time IPCR seemed to permit more convenient determination of specific serum IgE as well.
CONCLUSION
We believe that IPCR can serve as a valuable tool in determining specific serum IgE, especially when the amount of serum sample is limited.

Keyword

Blo t 5; Der p 1; Der p 2; IgE; allergy; immuno-PCR; real-time PCR

MeSH Terms

Allergens
Antigens, Dermatophagoides
Arthropod Proteins
Cysteine Endopeptidases
Enzyme-Linked Immunosorbent Assay
Humans
Hypersensitivity
Immunoassay
Immunoglobulin E
Limit of Detection
Mites
Polymerase Chain Reaction
Real-Time Polymerase Chain Reaction
Sensitivity and Specificity
Skin Tests
Allergens
Antigens, Dermatophagoides
Arthropod Proteins
Cysteine Endopeptidases
Immunoglobulin E
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