4.6 Article

A rapid magnetic particle-based enzyme immunoassay for human cytomegalovirus glycoprotein B quantification

期刊

出版社

ELSEVIER SCIENCE BV
DOI: 10.1016/j.jpba.2018.04.019

关键词

Magnetic beads; Immunoassay; mpEIA; Human cytomegalovirus; Glycoprotein B

资金

  1. FEDER funds through the POCI - COMPETE 2020 - Operational Program Competitiveness and Internationalization in Axis I - Strengthening research, technological development and innovation [POCI-01-0145-FEDRER-007491]
  2. Portuguese Funds by FCT - Foundation for Science and Technology [UID/Multi/00709/2013]
  3. Spanish Ministry of Science and Innovation (MICINN)
  4. Ministry of Economy and Competitiveness (MINECO)
  5. European Regional Development Fund (FEDER) [TEC20013-40561-P, MUSSEL RTC-2015-4077-2]

向作者/读者索取更多资源

Human cytomegalovirus (HCMV) is a herpes virus that can cause severe infections. Still, the available methods for its diagnostic have the main disadvantage of requiring long time to be performed. In this work, a simple magnetic particle-based enzyme immunoassay (mpEIA) for the quantification of glycoprotein B of Human cytomegalovirus (gB-HCMV) in urine samples is proposed. The immunosensor scheme is based on the analyte protein gB-HCMV sandwiched between a primary monoclonal antibody, (MBsPrG-mAbl), and a secondary anti-gB-HCMV antibody labelled with Horseradish peroxidase (Ab2-HRP) to allow spectrophotometric detection. The mpEIA analytical performance was tested in urine samples, showing a linear dependence between gB-HCMV concentration and the absorbance signal at 450 nm in a range of concentrations from 90 to 700 pg mL(-1). The calculated detection limits for gB-HCMV were 90 +/- 2 pg mL(-1) and the RSD was about 6.7% in urine samples. The immunosensor showed good selectivity against other viruses from Herpesviridae family, namely varicella zoster and Epstein Barr viruses. The recoveries of spiked human urine samples at 0.30-0.50 ng mL(-1) concentration levels of gB-HCMV ranged between 91 to 105%. The proposed mpEIA method was validated following the guidelines of the European Medicines Agency (EMEA-2014), and allows rapid, successful and easy quantification of gB-HCMV in urine samples. (C) 2018 Elsevier B.V. All rights reserved.

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