Development and Characterization of Double-Antibody Sandwich ELISA for Detection of Zika Virus Infection

Zhang, Liding; Du, Xuewei; Chen, Congjie; Zhi-xin, Chen; Zhang, Li; Han, Qinqin; Xia, Xueshan; Song, Yuzhu; Zhang, Jinyang

doi:10.3390/v10110634

Cited by 26 publications

(22 citation statements)

References 33 publications

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“…Recently, very few studies have been published for developing detection assays targeting ZIKV NS1 antigen. A double antibody sandwich ELISA-based colorimetric assay was able to detect it as low as 120 ng/mL [6]. Another sensing platform using a graphene biosensor was able to detect 500 ng/mL spiked in a 10-fold diluted serum [28].…”

Section: Resultsmentioning

confidence: 99%

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Development of a Flow-Free Automated Colorimetric Detection Assay Integrated with Smartphone for Zika NS1

Kabir

Zilouchian²,

Sher

et al. 2020

Diagnostics

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The Zika virus (ZIKV) is an emerging flavivirus transmitted to humans by Aedes mosquitoes that can potentially cause microcephaly, Guillain–Barré Syndrome, and other birth defects. Effective vaccines for Zika have not yet been developed. There is a necessity to establish an easily deployable, high-throughput, low-cost, and disposable point-of-care (POC) diagnostic platform for ZIKV infections. We report here an automated magnetic actuation platform suitable for a POC microfluidic sandwich enzyme-linked immunosorbent assay (ELISA) using antibody-coated superparamagnetic beads. The smartphone integrated immunoassay is developed for colorimetric detection of ZIKV nonstructural protein 1 (NS1) antigen using disposable chips to accommodate the reactions inside the chip in microliter volumes. An in-house-built magnetic actuator platform automatically moves the magnetic beads through different aqueous phases. The assay requires a total of 9 min to automatically control the post-capture washing, horseradish peroxidase (HRP) conjugated secondary antibody probing, washing again, and, finally, color development. By measuring the saturation intensity of the developed color from the smartphone captured video, the presented assay provides high sensitivity with a detection limit of 62.5 ng/mL in whole plasma. These results advocate a great promise that the platform would be useful for the POC diagnosis of Zika virus infection in patients and can be used in resource-limited settings.

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Section: Resultsmentioning

confidence: 99%

“…The nonstructural protein 1 (NS1) is targeted for detection due to its important role as a biomarker in flaviviruses. The protein is secreted by cells that contain the virus [6]. As the primary antigen, antibodies to NS1 can be formed in four to seven days, which can be used for detection.…”

Section: Introductionmentioning

confidence: 99%

Development of a Flow-Free Automated Colorimetric Detection Assay Integrated with Smartphone for Zika NS1

Kabir

Zilouchian²,

Sher

et al. 2020

Diagnostics

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“…Nonstructural protein 1 (NS1) is considered to be an essential biomarker for early detection and diagnosis of Zika virus. The development of double-antibody sandwich ELISA (DAS-ELISA) has been also used for early and rapid detection of ZIKA-NS1 protein in patients who newly infected with Zika virus [28].…”

Section: Clinical Subtypes Of Guillain-barre Syndrome (Gbs)mentioning

confidence: 99%

Guillain-Barre Syndrome and Miller Fisher Variant in Zika Virus Disease

Jadah¹

2021

Current Concepts in Zika Research

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Guillain-Barre syndrome (GBS) is a serious neurological disorder associated with a rapid progressive ascending muscle paralysis, and it is the most common neurological autoimmune disorder that affects the peripheral nervous system, which is usually triggered by viral or bacterial infection. GBS is rare in children and characterized by rapid progressive onset ascending muscle weakness associated with pain and sensory dysfunction. Miller Fisher syndrome (MFS), a variant of GBS, is rare in pediatric population which is typically manifested by ataxic gait, ophthalmoplegia, and areflexia since it is rare in children. It is vitally important to early diagnose this condition and to initiate early treatment to prevent further complications and long-term morbidity. Since the outbreak of Zika virus, the incidence of GBS has been increased. Zika virus associated with autoimmune anti-ganglioside antibodies trigger which lead to GBS development. Zika virus infection should be strongly considered in patients who present with classical signs of Miller Fisher syndrome, especially travelers and residents from endemic areas.

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“…Polyclonal antibodies (pAbs) and monoclonal antibodies (mAbs) against K. pneumoniae were produced as described by Zhang et al (24,25). Klebsiella pneumoniae was inactivated by treatment at 85°C for 1 hour, and then mixed with the Freund's complete adjuvant and injected into mice.…”

Section: Production Of Pabs and Mabs Against K Pneumoniaementioning

confidence: 99%

Visual and Rapid Detection of Klebsiella pneumoniae by Magnetic Immunocapture-Loop-Mediated Isothermal Amplification Assay

Zhang

Shi

Chen

et al. 2019

Jundishapur J Microbiol

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Background: Klebsiella pneumoniae is an important human pathogen that causes severe diseases including urinary tract infection, pneumonia, and bacteremia. However, a rapid and sensitive detection method remains to be developed. Objectives: This study aimed to develop a rapid, real-time, and visual detection method for K. pneumoniae. This is a primary screening method to improve the diagnosis of K. pneumoniae infection and save much precious time for clinical practice. Methods: Klebsiella pneumoniae was used as an antigen to produce a monoclonal antibodies (mAb). The mAb 1E6 recognizing outer membrane protein C on the surface of K. pneumoniae was screened by an indirect enzyme-linked immunosorbent assay (ELISA), Western Blot, and mass spectrometry assay, and then conjugated with the protein A/G coated magnetic beads to generate the immune magnetic beads (IMBs). Thereafter, the IMBs were used to capture K. pneumoniae, and the complex (beads-mAb-K. pneumoniae) was used for loop-mediated isothermal amplification (LAMP) assay. Results: We developed a rapid, real-time, and visual detection method employing an immunocapture loop-mediated isothermal amplification (IC-LAMP). The sensitivity of the IC-LAMP was 4 CFU mL-1. The process of K. pneumoniae detection lasted~60 minutes and had no cross-reaction to other microbial strains. Besides, the accuracy of IC-LAMP was further verified by examining 39 clinical isolates. Conclusions: Without the need for enrichment of bacteria and extraction of its genome, the IC-LAMP developed here could be used as a primary screening method supplementary to traditional detection methods to improve the diagnosis of K. pneumoniae infection for clinical practice.

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Development and Characterization of Double-Antibody Sandwich ELISA for Detection of Zika Virus Infection

Cited by 26 publications

References 33 publications

Development of a Flow-Free Automated Colorimetric Detection Assay Integrated with Smartphone for Zika NS1

Development of a Flow-Free Automated Colorimetric Detection Assay Integrated with Smartphone for Zika NS1

Guillain-Barre Syndrome and Miller Fisher Variant in Zika Virus Disease

Visual and Rapid Detection of Klebsiella pneumoniae by Magnetic Immunocapture-Loop-Mediated Isothermal Amplification Assay

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