Basic Science Investigation
Establishment and performance evaluation of Der p 1/Der p 2 specific immunoglobulin E detection for the components of dust mite allergens based on chemiluminescence immunoassay
Han Feifei, Zhou Ying, Zhang Jian, Yin Hao, Wu Meili, Chen Xi, Cui Yubao
Published 2022-03-25
Cite as Chin J Nucl Med Mol Imaging, 2022, 42(3): 160-164. DOI: 10.3760/cma.j.cn321828-20200817-00314
Abstract
ObjectiveTo establish a quantitative detection method for the main components of dust mite allergens Der p 1, Der p 2 specific immunoglobulin E (sIgE) by using the nano-magnetic particle chemiluminescence immunoassay.
MethodsThe performance indexes of the established method were evaluated after setting up and optimizing the chemiluminescence detection system and immune reaction conditions of sIgE for dust mite allergen. Serum sIgE levels of 50 suspected allergic patients with dust mite were determined by this chemiluminescence method. At the same time, this method was compared with the Phadia kit and the consistency was analyzed by Kappa test.
ResultsThe optimal amount of magnetic beads was 25 μg, the optimal reaction buffer (pH=7.4) contained 0.1 mol/L Tris-HCl and 0.25%(W/W) casein, the optimal coating solution contatined 20 mmol/L phosphate buffer (PB) and 1%(W/W) bovine serum albumin (BSA), and the luminescence enhancement solution contained 0.05%(V/V) Triton X-100. The two-step immunoreaction was adopted, and the detection could be completed with 20 μl sample at the optimal reaction temperature of 37℃. The limit of detection (LOD) of the established nano-magnetic particle chemiluminescence system in detecting Der p 1 and Der p 2 sIgE antibodies were both less than 0.01 kU/L, with the linear range of 0.2-100.0 kU/L, the precision of less than 7%, and the cross contamination rate of 0.19% and 0.21%. Compared with the Phadia system, the positive and negative coincidence rate of Der p 1 were 78.0%(32/41) and 9/9 with good consistency (Kappa=0.65, P=0.008), and the positive and negative coincidence rate of Der P 2 were 93.3%(28/30) and 85.0%(17/20) with good consistency (Kappa=0.79, P=0.003).
ConclusionThe nano-magnetic particle chemiluminescence immunoassay is successfully established for detecting dust mite allergen sIgE, which has good detection performance and good consistency with Phadia system.
Key words:
Immunoglobulin E; Antigens, dermatophagoides; Dermatophagoides pteronyssinus; Chemiluminescent measurements; Nanoparticles
Contributor Information
Han Feifei
Department of Medical Laboratory, Wuxi People′s Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China
Zhou Ying
Pediatric Laboratory, Wuxi Children′s Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China
Zhang Jian
Department of Medical Laboratory, Wuxi Children′s Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China
Yin Hao
Department of Medical Laboratory, Wuxi People′s Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China
Wu Meili
Pediatric Laboratory, Wuxi Children′s Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China
Chen Xi
Department of Medical Laboratory, Wuxi Children′s Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China
Cui Yubao
Department of Medical Laboratory, Wuxi People′s Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China
