Disposable immunosensor for human cytomegalovirus glycoprotein B detection

Pires, Filipe; Silva, H.; Domínguez‐Renedo, Olga; Alonso‐Lomillo, M. Asunción; Arcos‐Martínez, M. Julia; Dias-Cabral, A.C.

doi:10.1016/j.talanta.2014.12.016

Cited by 10 publications

(14 citation statements)

References 19 publications

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“…This is quite advantageous since the collection of the biological samples is simple and is performed by noninvasive procedures [7]. In consequence, the development of methods for the accurate detection of gB-HCMV in body fluids, like urine and saliva, where viral loads are higher, is of great interest, unlike the up mentioned analytical method based on the determination of antibodies generated by the HCMV virus [6,13].…”

Section: Accepted Manuscriptmentioning

confidence: 99%

“…We have previously reported an electrochemical immunosensor for gB-HCMV detection, by using capture anti-gB-HCMV antibodies absorbed on screen-printed carbon electrodes, and secondary anti-gB-HCMV antibodies labelled with gold nanoparticles [6]. gB-HCMV detection was carried out through electrochemical stripping analysis of silver nanoparticles deposited quantitatively on the immunosensor, catalyzed by the nanogold labels [6].…”

Section: Accepted Manuscriptmentioning

confidence: 99%

“…The reactivation of HCMV to a state of active replication with potential to induce disease and transmission to a new host may occur in situations of immune system dysfunction or low maturity [1,2]. Severe clinical symptoms associated with HCMV primary infection or reactivation are observed in immunosuppressed individuals, like people who were transplanted, infected by human immunodeficiency virus, or with an immature immune system, like fetus and newborns infants [4,5,6].…”

Section: Introductionmentioning

confidence: 99%

“…The traditional method for direct free HCMV detection is viral isolation, that detect viral antigens in tissue, urine or saliva samples [6,10]. Nevertheless, time analysis requirements for this procedure (≈ 20 days before the sample can be reported as negative) makes it disadvantageous to be routinely used as point of care test in clinical diagnosis [6,10].…”

Section: Introductionmentioning

confidence: 99%

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

Pires

Arcos‐Martínez

Dias-Cabral

et al. 2018

Journal of Pharmaceutical and Biomedical Analysis

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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, (MBs-PrG-mAb1), 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. The calculated detection limits for gB-HCMV were 90 ± 2 pg mL 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 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.

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Section: Accepted Manuscriptmentioning

confidence: 99%

Section: Accepted Manuscriptmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

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

Pires

Arcos‐Martínez

Dias-Cabral

et al. 2018

Journal of Pharmaceutical and Biomedical Analysis

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“…Au nanoparticle-labeled antibodies can be used as catalysts in electrochemical stripping analysis. Au nanoparticle-based immunosensors are prepared by modifying the surface of a screen-printed electrode with anti-human cytomegalovirus glycoprotein B (anti-human cytomegalovirus (HCMV) glycoprotein B) [33]. The antibody-modified electrode is further incubated in solutions of HCMV glycoprotein B and anti-HCMV glycoprotein B labeled with Au nanoparticles.…”

Section: Immunosensors For Glycoproteinsmentioning

confidence: 99%

Recent Progress in Electrochemical Biosensors for Glycoproteins

Akiba

Anzai

2016

Sensors

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This review provides an overview of recent progress in the development of electrochemical biosensors for glycoproteins. Electrochemical glycoprotein sensors are constructed by combining metal and carbon electrodes with glycoprotein-selective binding elements including antibodies, lectin, phenylboronic acid and molecularly imprinted polymers. A recent trend in the preparation of glycoprotein sensors is the successful use of nanomaterials such as graphene, carbon nanotube, and metal nanoparticles. These nanomaterials are extremely useful for improving the sensitivity of glycoprotein sensors. This review focuses mainly on the protocols for the preparation of glycoprotein sensors and the materials used. Recent improvements in glycoprotein sensors are discussed by grouping the sensors into several categories based on the materials used as recognition elements.

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Electrochemical Immunoassay for Cytomegalovirus Antigen Detection with Multiple Signal Amplification Using HRP and Pt‐Pd Nanoparticles Functionalized Single‐walled Carbon Nanohorns

et al. 2016

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Cytomegalovirus is typically associated with immunocompromised hosts, pregnant women and transplant patients, who require a timely diagnosis. In this work, a sensitive and highly specific electrochemical amplification immunosensor was established for detecting Cytomegalovirus pp65 antigen based on Pt‐PdNPs@SWCNHs with horseradish peroxidase (HRP) as a signal enhancer and thionine as a signal probe. First, Pt nanoparticle (PtNP) and Pd nanoparticle (PdNP) functionalized single‐walled carbon nanohorn (SWCNH) nanocomposites, i.e. Pt‐PdNPs@SWCNHs, was used as a carrier for immobilization of antibody through the Pt‐N bond and the Pd‐N bond. Next, HRP was used to block the rest of the binding‐sites. Signal amplification was obtained by the cooperative catalytic activities of Pt‐PdNPs and HRP to H2O2. SWCNHs loaded with a large amount of Pt‐PdNPs further amplified the signal due to the excellent surface area. The fabricated immunosensor was used to detect different concentrations of Cytomegalovirus pp65 antigen under optimized conditions. The tests showed a linear range from 0.1 to 80 ng mL−1 with a low detection limit of 30 pg mL−1, and exhibited excellent selectivity, stability and reproducibility. Therefore, this project presented a potential approach for the early diagnosis of Cytomegalovirus infection in clinical trials.

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Disposable immunosensor for human cytomegalovirus glycoprotein B detection

Cited by 10 publications

References 19 publications

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

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

Recent Progress in Electrochemical Biosensors for Glycoproteins

Electrochemical Immunoassay for Cytomegalovirus Antigen Detection with Multiple Signal Amplification Using HRP and Pt‐Pd Nanoparticles Functionalized Single‐walled Carbon Nanohorns

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