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. 2022 Sep 1;23(17):9991.
doi: 10.3390/ijms23179991.

Lateral Flow Immunoassay Based on Time-Resolved Fluorescence Microspheres for Rapid and Quantitative Screening CA199 in Human Serum

Xueshima Jiao  1   2 , Tao Peng  2 , Zhanwei Liang  1   2 , Yalin Hu  1   2 , Bo Meng  2 , Yang Zhao  2 , Jie Xie  2 , Xiaoyun Gong  2 , You Jiang  2 , Xiang Fang  2 , Xiaoping Yu  1 , Xinhua Dai  2
Affiliations

Lateral Flow Immunoassay Based on Time-Resolved Fluorescence Microspheres for Rapid and Quantitative Screening CA199 in Human Serum

Xueshima Jiao et al. Int J Mol Sci. .

Abstract

Carbohydrate antigen 199 (CA199) is a serum biomarker which has certain value and significance in the diagnosis, prognosis, treatment, and postoperative monitoring of cancer. In this study, a lateral flow immunoassay based on europium (III) polystyrene time-resolved fluorescence microspheres (TRFM-based LFIA), integrated with a portable fluorescence reader, has been successfully establish for rapid and quantitative analysis of CA199 in human serum. Briefly, time-resolved fluorescence microspheres (TRFMs) were conjugated with antibody I (Ab1) against CA199 as detection probes, and antibody II (Ab2) was coated as capture element, and a "TRFMs-Ab1-CA199-Ab2" sandwich format would form when CA199 was detected by the TRFM-based LFIA. Under the optimal parameters, the detection limit of the TRFM-based LFIA for visible quantitation with the help of an ultraviolet light was 4.125 U/mL, which was four times lower than that of LFIA based on gold nanoparticles. Additionally, the fluorescence ratio is well linearly correlated with the CA199 concentration (0.00-66.0 U/mL) and logarithmic concentration (66.0-264.0 U/mL) for quantitative detection. Serum samples from 10 healthy people and 10 liver cancer patients were tested to confirm the performances of the point-of-care application of the TRFM-based LFIA, 20.0 U/mL of CA199 in human serum was defined as the threshold for distinguishing healthy people from liver cancer patients with an accuracy of about 60%. The establishment of TRFM-based LFIA will provide a sensitive, convenient, and efficient technical support for rapid screening of CA199 in cancer diagnosis and prognosis.

Keywords: biomarker; cancer; carbohydrate antigen 199; lateral flow immunoassay; time-resolved fluorescent microspheres.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Schematic of the TRFM-based LFIA (A) for CA199 rapid qualitative (B) and quantitative (C) detection.
Figure 2
Figure 2
Qualitative and quantitative detection results of the TRFM-based LFIA for CA199. Visible results (A) and FIt/c values (B) of TRFM-based LFIA with different concentrations of CA199. The linear ranges for quantitatively detecting CA199 were constructed by plotting the fluorescence ratio versus the concentration 0.00–66.0 U/mL (C) and the logarithmic concentration 66.0–264.0 U/mL (D), respectively.
Figure 3
Figure 3
Visible result (A) and FIt/c (B) value of the TRFM-based LFIA for specificity evaluation.
Figure 4
Figure 4
Qualitative results of CA199 in liver cancer patients (A) and healthy people (B), CA199 levels quantitatively detected by TRFM-based LFIA (C).
Figure 5
Figure 5
Performances of the AuNP-based LFIA for CA199. (A) Comparison between TRFM-based LFIA and AuNP-based LFIA in testing CA199 from 0.00 U/mL to 528.0 U/mL. (B) T/C ratio of CA199 detected by the AuNP-based LFIA. (C,D) Calibration curves for the determination of CA199.
See this image and copyright information in PMC

References

    1. Navaneethan U., Lourdusamy V., Poptic E., Hammel J.P., Sanaka M.R., Parsi M.A. Comparative effectiveness of pyruvate kinase M2 in bile, serum carbohydrate antigen 19-9, and biliary brushings in diagnosing malignant biliary strictures. Dig. Dis. Sci. 2015;60:903–909. doi: 10.1007/s10620-014-3397-7. - DOI - PubMed
    1. Wang Y., Mi Y., Li Y., Wang J., Cheng L., Yan S., Deng L. Establishment of time-resolved fluorescence immunochromatographic assay for detection of carbohydrate antigen 19-9. Chin. J. Biotechnol. 2018;34:1012–1018. doi: 10.13345/j.cjb.170470. - DOI - PubMed
    1. Luo G., Jin K., Deng S., Cheng H., Fan Z., Gong Y., Qian Y., Huang Q., Ni Q., Liu C., et al. Roles of CA19-9 in pancreatic cancer: Biomarker, predictor and promoter. Biochim. Et Biophys. Acta Rev. Cancer. 2021;1875:188409. doi: 10.1016/j.bbcan.2020.188409. - DOI - PubMed
    1. Qian L., Li Q., Baryeh K., Qiu W., Li K., Zhang J., Yu Q., Xu D., Liu W., Brand R.E., et al. Biosensors for early diagnosis of pancreatic cancer: A review. Transl. Res. 2019;213:67–89. doi: 10.1016/j.trsl.2019.08.002. - DOI - PubMed
    1. Liang Y., Wang W., Fang C. Clinical significance and diagnostic value of serum CEA, CA19-9 and CA72-4 in patients with gastric cancer. Oncotarget. 2016;7:49565–49573. doi: 10.18632/oncotarget.10391. - DOI - PMC - PubMed