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

作者：Pires F; Julia Arcos Martinez M; Cristina Dias Cabral A; Vidal Juan C; Castillo Juan R

来源：Journal of Pharmaceutical and Biomedical Analysis, 2018, 156: 372-378.

DOI：10.1016/j.jpba.2018.04.019

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

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.

• 出版日期2018-7-15