A dual‐label time‐resolved fluorescence immunoassay for the simultaneous determination of ferritin and β<sub>2</sub>‐microglobulin

Liu, Zhi; Huang, Jing; Ou, Ruiming; Yao, Mengdong; She, Yan-lin; Chen, Rui; Li, Cheng; Li, Xu; Abudureyimu, Aikebaier; Zhang, Qing; Liu, Shuang

doi:10.1002/jcla.22132

Cited by 8 publications

(6 citation statements)

References 16 publications

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“…Due to the high sensitivity and specificity, dual-marker and multi-marker testing is necessary for clinical practice. For example, the simultaneous determinaton of ferritin and β2‐microglobulin could be used for the early screening and follow-up surveillance of the acute and chronic LL [ 23 ]. Comparison results of the concentrations of S100A8 and LRG1 in 120 healthy serum samples and 59 serum samples with LL, the concentrations of S100A8 and LRG1 in the LL patients was significantly higher than that of healthy human.…”

Section: Discussionmentioning

confidence: 99%

A new method for screening acute/chronic lymphocytic leukemia: dual-label time-resolved fluorescence immunoassay

Zhang

Dai

et al. 2022

BMC Biotechnol

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Background Lymphocytic leukemia (LL) is a primary malignant tumor of hematopoietic tissue, which seriously affects the health of children and the elderly. The study aims to establish a new detection method for screening acute/chronic LL using time-resolved fluorescence immunoassay (TRFIA) via quantitative detection of S100 calcium binding protein A8 (S100A8) and leucine-rich alpha-2-glycoprotein 1 (LRG1) in serum. Methods Here a sandwich TRFIA was optimized and established: Anti-S100A8/LRG1 caputre antibodies immobilized on 96-well plates captured S100A8/LRG1, and then banded together with the anti-S100A8/LRG1 detection antibodies labeled with Europium(III) (Eu3+)/samarium(III) (Sm3+) chelates. Finally time resolved fluorometry measured the fluorescence intensity. Results The sensitivity of S100A8 was 1.15 ng/mL(LogY = 3.4027 + 0.4091 × LogX, R2 = 0.9828, P < 0.001, dynamic range: 2.1–10,000 ng/mL), and 3.2 ng/mL for LRG1 (LogY = 3.3009 + 0.4082 × LogX, R2 = 0.9748, P < 0.001, dynamic range: 4.0–10,000 ng/mL). The intra-assay and inter-assay CVs were low, ranging from 5.75% to 8.23% for S100A8 and 5.30% to 9.45% for LRG1 with high specificity and affinity in serum samples. Bland–Altman plots indicated TRFIA and ELISA kits have good agreement in clinical serum samples. Additionally, the cutoff values for S100A8 and LRG1 were 1849.18 ng/mL and 588.08 ng/mL, respectively. Conclusion The present TRFIA method could be used for the quantitative detection of S100A8 and LRG1 in serum, and it has high sensitivity, accuracy and specificity. Clinically, this TRFIA method could be suitable for screening of LL via the quantitative detection of S100A8 and LRG1.

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Section: Discussionmentioning

confidence: 99%

A new method for screening acute/chronic lymphocytic leukemia: dual-label time-resolved fluorescence immunoassay

Zhang

Dai

et al. 2022

BMC Biotechnol

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“…Due to different emission wavelengths and kinetics, lanthanide chelates (Sm 3+ , Eu 3+ , Tb 3+ , etc.) have no nonspecific background (interferents), which results in the TRFIA showing better performance in antibody detection in serum [12,13]. Compared with the colloidal gold method and chemiluminescence method, the TRFIA method has a higher sensitivity and wide dynamic range [14].…”

Section: Discussionmentioning

confidence: 99%

“…have no nonspecific background (interferents), which results in the TRFIA showing better performance in antibody detection in serum. 12 , 13 Compared with the colloidal gold method and chemiluminescence method, the TRFIA method has a higher sensitivity and wide dynamic range. 14 Importantly, the double‐antigen sandwich method has higher sensitivity than the double‐antibody sandwich method and indirect ELISA method.…”

Section: Discussionmentioning

confidence: 99%

Determination of human COVID‐19 total antibodies in serum using a time‐resolved fluorescence immunoassay

Chen

Lai

et al. 2021

Biotech and App Biochem

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Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is spreading rapidly around the world. Antibody detection This article is protected by copyright. All rights reserved.an important role in the diagnosis of COVID-19. Here, we established a new time-resolved fluorescence immunoassay (TRFIA) to determine COVID-19 total antibodies. A double antigen sandwich TRFIA was optimized and established: recombinant nucleocapsid phosphoprotein (N protein) and spike protein (S protein) of COVID-19 immobilized on 96-well plates captured human COVID-19 antibodies and then banded together with the N/S proteins labeled with europium(III) (Eu 3+ ) chelates, and finally, time-resolved fluorometry was used to measure the fluorescence values. We successfully established a TRFIA method for the detection of human COVID-19 total antibodies, and the cutoff value was 2.02. There was no cross-reactivity with the negative reference of the National Reference Panel for IgM and IgG antibodies to COVID-19. The CV of the precision assay was 3.19%, and the assay could be stored stably for 15 days at 37°C. Compared with that of the colloidal gold method and chemiluminescence method, the sensitivity of the TRFIA method was higher, and the false positive/negative rate was lower. This established TRFIA has high sensitivity, accuracy and specificity, which indicates that this method provides a new detection method for the high-throughput routine diagnosis of COVID-19.

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“…In TRFIA detection, the hook effect appears at high substance concentration [8], which makes TRFIA detection has a wide dynamic range. In a word, TRFIA has high sensitivity and wide dynamic range [23,24], and it might be a promising detection technique for quantitative detection of CPV-2-Ag in vaccine.…”

Section: Figmentioning

confidence: 99%

Determination of parvovirus antigen in the vaccine using time‐resolved fluorescence immunoassay

Chen¹,

Hang²,

Hu³

et al. 2020

Biotech and App Biochem

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As a highly contagious and potentially fatal disease of dogs, canine parvovirus type 2 (CPV-2) usually causes severe myocarditis and gastroenteritis, while vaccine injection has greatly reduced the incidence of CPV-2 diseases. However, there is currently a lack of simple and effective method for quantitative detection of CPV-2 in vaccine. Therefore, this study aims to prepare an accurate method to determine the CPV-2 antigen (CPV-2-Ag) in vaccine. Here, a sandwich time-resolved fluorescence immunoassay (TRFIA) was established and optimized. Anti-CPV-2 antibodies were immobilized on 96-well plates to capture CPV-2-Ag, and then bound together with the detection antibodies labeled with Europium(III) (Eu 3+ ) chelates; finally, time-resolved fluorometry was employed to measure the fluorescence intensity. Vaccination was performed to evaluate the relationship between CPV-2-Ag concentration and antibody titer. The sensitivity is 1.15 mEU/mL (LogY = 1.524 + 0.8667 × LogX, R 2 = 0.9933), and the average recovery is among 91.00% to 106.39% without cross-reactions with the other canine viral antigen. The correlation between ELISA assay and this method is up to 0.9861. And, there is high correlation between the CPV-2-Ag concentration and antibody titers (R 2 = 0.9234). This immunoassay established has high sensitivity, accuracy, and specificity, which indicate that this method could be suitable for quantitative detection of CPV-2-Ag in vaccine evaluation.

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A dual‐label time‐resolved fluorescence immunoassay for the simultaneous determination of ferritin and β₂‐microglobulin

Abstract: The present dual-label TRFIA has high sensitivity, specificity, and accuracy in clinical sample analysis. It is an effective detection method for the early screening and follow-up surveillance of the acute and chronic lymphocytic leukemia.

Cited by 8 publications

References 16 publications

A new method for screening acute/chronic lymphocytic leukemia: dual-label time-resolved fluorescence immunoassay

A new method for screening acute/chronic lymphocytic leukemia: dual-label time-resolved fluorescence immunoassay

Determination of human COVID‐19 total antibodies in serum using a time‐resolved fluorescence immunoassay

Determination of parvovirus antigen in the vaccine using time‐resolved fluorescence immunoassay

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A dual‐label time‐resolved fluorescence immunoassay for the simultaneous determination of ferritin and β2‐microglobulin

Abstract: The present dual-label TRFIA has high sensitivity, specificity, and accuracy in clinical sample analysis. It is an effective detection method for the early screening and follow-up surveillance of the acute and chronic lymphocytic leukemia.

Cited by 8 publications

References 16 publications

A new method for screening acute/chronic lymphocytic leukemia: dual-label time-resolved fluorescence immunoassay

A new method for screening acute/chronic lymphocytic leukemia: dual-label time-resolved fluorescence immunoassay

Determination of human COVID‐19 total antibodies in serum using a time‐resolved fluorescence immunoassay

Determination of parvovirus antigen in the vaccine using time‐resolved fluorescence immunoassay

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Product

Resources

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A dual‐label time‐resolved fluorescence immunoassay for the simultaneous determination of ferritin and β₂‐microglobulin