Development of a fluorescent immnunochromatographic assay for the procalcitonin detection of clinical patients in China.

Abstract

BACKGROUND:

Procalcitonin (PCT) has been recognized as a biomarker in severe inflammation, infection and sepsis. PCT detection in serum requires sensitive and specific antibodies. In this study, we generated monoclonal antibodies (mAbs) and developed fluorescent immunochromatographic assay for PCT detection.

METHODS:

Human recombinant PCT was used as immunogen. mAbs against PCT were developed and applied to fluorescent immunochromatographic assay for PCT detection in clinical samples.

RESULTS:

Out of 35 hybridoma cell lines secreting antibodies against the recombinant PCT, five sensitive and specific cell lines were selected and designated as F6, G2, C2, D2 and E5. All these antibodies have no cross reaction with calcitonin or calcitonin gene-related peptides (CGRP). After screening for pairing, mAb F6 was labeled with fluorescent microspheres and C2 was coated on a nitrocellulose membrane for immunochromatographic test. All 35 clinical samples were detected by the mAb F6-C2 test strips and the bioMérieux PCT assay. The test strips showed high specificity and sensitivity for PCT. Good correlation was observed between our immunochromatographic test strips and the bioMérieux PCT assay (R(2):0.986).

CONCLUSIONS:

These newly developed anti-PCT mAbs and fluorescent immunochromatographic assay can serve as important diagnostic tools for a fast, reliable and point-of-care testing for easy determination of PCT in serum and diagnosis of bacterial infection, inflammation or sepsis.

• Authors: Haiying Wang, Hong Wang, Shaopei Chen, Emmanuel E Dzakah, Keren Kang, Jihua Wang, Jufang Wang
• Journal: Clinica chimica acta; international journal of clinical chemistry (Clin Chim Acta) Vol. 444 Pg. 37-42 (Apr 15 2015) ISSN: 1873-3492 [Electronic] Netherlands
• PMID: 25659293 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2015 Elsevier B.V. All rights reserved.

Chemical References

• Antibodies, Monoclonal
• CALCA protein, human
• Calca protein, mouse
• Protein Precursors
• Recombinant Proteins
• Calcitonin
• Calcitonin Gene-Related Peptide

Topics

• Animals
• Antibodies, Monoclonal (immunology)
• Antigen-Antibody Reactions
• Calcitonin (blood, genetics, immunology)
• Calcitonin Gene-Related Peptide
• China
• Chromatography, Affinity (methods)
• Enzyme-Linked Immunosorbent Assay
• Female
• Fluorescence
• Humans
• Mice
• Mice, Inbred BALB C
• Protein Precursors (blood, genetics, immunology)
• Recombinant Proteins (blood, genetics, immunology)