Multiplex Microsphere Immunoassay Identifies Three Flavivirus Infections
|
By LabMedica International staff writers Posted on 19 Sep 2019 |
|
Image: The MagPlex Multiplexing Microspheres (Photo courtesy of Luminex).
The explosive spread of Zika virus (ZIKV) and associated complications in flavivirus-endemic regions underscore the need for sensitive and specific serodiagnostic tests to distinguish ZIKV, dengue virus (DENV) and other flavivirus infections.
ZIKV is a member of the genus Flavivirus of the family Flaviviridae, which includes several pathogenic mosquito-borne viruses in different serocomplexes. The four serotypes of dengue virus (DENV) belong to the DENV serocomplex; West Nile virus (WNV) and Japanese encephalitis virus (JEV) to the JEV serocomplex; yellow fever virus (YFV) as a single member; and ZIKV.
Tropical medicine specialists at the University of Hawaii at Manoa, (Honolulu, HI, USA) and their colleagues developed a high-throughput and multiplex IgG microsphere immunoassay (MIA) using the NS1 proteins of DENV1-DENV4, ZIKV and West Nile virus (WNV) to test samples from reverse-transcription-polymerase-chain reaction-confirmed cases, including primary DENV1, DENV2, DENV3, WNV and ZIKV infections, secondary DENV infection, and ZIKV infection with previous DENV infection.
The NS1 gene (corresponding to amino acid residues 1–352) of ZIKV (HPF2013 strain) with a His-tag at the C-terminus was codon-optimized. Six purified NS1 proteins, bovine serum albumin (BSA) and PBS (as negative antigen control) were coupled individually onto eight types of magnetic carboxylated microsphere beads containing different fluorophores (MagPlexTM-C) using two-step carbodiimide process at room temperature. Microsphere immunoassays (MIA) were performed and read by a Luminex 200 machine. Enzyme-linked immunosorbent assays were performed for DENV1-, DENV2-, DENV3-, and ZIKV-NS1 IgG.
The scientists reported that a combination of four DENV NS1 IgG MIAs revealed a sensitivity of 94.3% and specificity of 97.2% to detect DENV infection. The ZIKV and WNV NS1 IgG MIAs had a sensitivity/specificity of 100%/87.9% and 86.1%/78.4%, respectively. A positive correlation was found between the readouts of ELISA and MIA for the different NS1 tested. Based on the ratio of relative median fluorescence intensity of ZIKV NS1 to DENV1 NS1, the IgG MIA can distinguish ZIKV infection with previous DENV infection and secondary DENV infection with a sensitivity of 88.9%–90.0% and specificity of 91.7%–100.0%.
The authors concluded that the multiplex and high-throughput assay could be applied to serodiagnosis and serosurveillance of DENV, ZIKV and WNV infections in endemic regions. The study was published on August 23, 2019, in the journal PLOS Neglected Tropical Diseases.
Related Links:
University of Hawaii at Manoa
ZIKV is a member of the genus Flavivirus of the family Flaviviridae, which includes several pathogenic mosquito-borne viruses in different serocomplexes. The four serotypes of dengue virus (DENV) belong to the DENV serocomplex; West Nile virus (WNV) and Japanese encephalitis virus (JEV) to the JEV serocomplex; yellow fever virus (YFV) as a single member; and ZIKV.
Tropical medicine specialists at the University of Hawaii at Manoa, (Honolulu, HI, USA) and their colleagues developed a high-throughput and multiplex IgG microsphere immunoassay (MIA) using the NS1 proteins of DENV1-DENV4, ZIKV and West Nile virus (WNV) to test samples from reverse-transcription-polymerase-chain reaction-confirmed cases, including primary DENV1, DENV2, DENV3, WNV and ZIKV infections, secondary DENV infection, and ZIKV infection with previous DENV infection.
The NS1 gene (corresponding to amino acid residues 1–352) of ZIKV (HPF2013 strain) with a His-tag at the C-terminus was codon-optimized. Six purified NS1 proteins, bovine serum albumin (BSA) and PBS (as negative antigen control) were coupled individually onto eight types of magnetic carboxylated microsphere beads containing different fluorophores (MagPlexTM-C) using two-step carbodiimide process at room temperature. Microsphere immunoassays (MIA) were performed and read by a Luminex 200 machine. Enzyme-linked immunosorbent assays were performed for DENV1-, DENV2-, DENV3-, and ZIKV-NS1 IgG.
The scientists reported that a combination of four DENV NS1 IgG MIAs revealed a sensitivity of 94.3% and specificity of 97.2% to detect DENV infection. The ZIKV and WNV NS1 IgG MIAs had a sensitivity/specificity of 100%/87.9% and 86.1%/78.4%, respectively. A positive correlation was found between the readouts of ELISA and MIA for the different NS1 tested. Based on the ratio of relative median fluorescence intensity of ZIKV NS1 to DENV1 NS1, the IgG MIA can distinguish ZIKV infection with previous DENV infection and secondary DENV infection with a sensitivity of 88.9%–90.0% and specificity of 91.7%–100.0%.
The authors concluded that the multiplex and high-throughput assay could be applied to serodiagnosis and serosurveillance of DENV, ZIKV and WNV infections in endemic regions. The study was published on August 23, 2019, in the journal PLOS Neglected Tropical Diseases.
Related Links:
University of Hawaii at Manoa
Gold Member
Quantitative POC Immunoassay Analyzer
EASY READER+
New
Pneumonia Test
Pneumocystis ELITe MGB® Kit
New
Rubella Virus Test
RUB IgM ELISA
Latest Immunology News
- Microfluidic Chip Detects Cancer Recurrence from Immune Response Signals
- Cancer Mutation ‘Fingerprints’ to Improve Prediction of Immunotherapy Response
- Immune Signature Identified in Treatment-Resistant Myasthenia Gravis
- New Biomarker Predicts Chemotherapy Response in Triple-Negative Breast Cancer
- Blood Test Identifies Lung Cancer Patients Who Can Benefit from Immunotherapy Drug
- Whole-Genome Sequencing Approach Identifies Cancer Patients Benefitting From PARP-Inhibitor Treatment
- Ultrasensitive Liquid Biopsy Demonstrates Efficacy in Predicting Immunotherapy Response
- Blood Test Could Identify Colon Cancer Patients to Benefit from NSAIDs
- Blood Test Could Detect Adverse Immunotherapy Effects
- Routine Blood Test Can Predict Who Benefits Most from CAR T-Cell Therapy
- New Test Distinguishes Vaccine-Induced False Positives from Active HIV Infection
- Gene Signature Test Predicts Response to Key Breast Cancer Treatment
- Chip Captures Cancer Cells from Blood to Help Select Right Breast Cancer Treatment
- Blood-Based Liquid Biopsy Model Analyzes Immunotherapy Effectiveness
- Signature Genes Predict T-Cell Expansion in Cancer Immunotherapy
- Molecular Microscope Diagnostic System Assesses Lung Transplant Rejection
Channels
Clinical Chemistry
view channel
New Plasma Tau Assay Improves Prediction of Alzheimer’s Progression
Predicting which patients with early symptomatic Alzheimer’s disease will decline more rapidly remains a key challenge in both research and patient care. Growing interest in tau biology, along with advances... Read more
Routine Blood Markers Predict Heart Failure Risk in Prediabetes
Heart failure prevention relies on finding high-risk adults before symptoms appear, yet effective stratification remains difficult in routine care. Prediabetes affects an estimated 115.2 million U.... Read more
Molecular Diagnostics
view channel
Blood Biomarker Predicts Cognitive Outcomes After Cardiac Arrest
Long-term cognitive impairment is a frequent consequence of out-of-hospital cardiac arrest yet early prediction remains difficult. Clinicians commonly use blood-based markers to estimate brain injury risk... Read more
Liquid Biopsy Enables Faster Diagnosis of Childhood Cancer in Africa
Burkitt lymphoma is the most common childhood cancer in Africa and progresses rapidly, making fast, accurate diagnosis essential to survival. Although survival can exceed 90% when therapy starts quickly,... Read more
Hematology
view channel
Rapid Cartridge-Based Test Aims to Expand Access to Hemoglobin Disorder Diagnosis
Sickle cell disease and beta thalassemia are hemoglobin disorders that often require referral to specialized laboratories for definitive diagnosis, delaying results for patients and clinicians.... Read more
New Guidelines Aim to Improve AL Amyloidosis Diagnosis
Light chain (AL) amyloidosis is a rare, life-threatening bone marrow disorder in which abnormal amyloid proteins accumulate in organs. Approximately 3,260 people in the United States are diagnosed... Read more
Immunology
view channel
Microfluidic Chip Detects Cancer Recurrence from Immune Response Signals
Early identification of treatment response and relapse remains a major challenge in solid tumors, where minimal residual disease is difficult to detect with routine imaging and blood tests.... Read more
Cancer Mutation ‘Fingerprints’ to Improve Prediction of Immunotherapy Response
Cancer cells accumulate thousands of genetic mutations, but not all mutations affect tumors in the same way. Some make cancer cells more visible to the immune system, while others allow tumors to evade... Read more
Pathology
view channel
AI-Powered Tool to Transform Dermatopathology Workflow
Skin cancer accounts for the largest number of cancer diagnoses in the United States, placing sustained pressure on pathology services. Diagnostic interpretation can be variable for challenging melanocytic... Read more
New Chromogenic Culture Media Enable Rapid Detection of Candida Infections
Invasive Candida infections are challenging for healthcare systems, with some strains spreading rapidly in hospitals and showing resistance to multiple antifungal drugs. Candida auris is associated with... Read more
Technology
view channel
Portable Breath Sensor Detects Pneumonia Biomarkers in Minutes
Pneumonia is commonly confirmed with chest X-rays or laboratory assays that can take hours, delaying clinical decisions in acute and outpatient settings. Breath-based diagnostics promise faster answers... Read more
New Electronic Pipette Enhances Workflows with Touchscreen Control
Manual pipetting remains a routine yet error-prone step that can affect reproducibility and throughput in clinical and research laboratories. Training demands and ergonomic strain also add variability... Read more
AI Model Outperforms Clinicians in Rare Disease Detection
Rare diseases affect an estimated 300 million people worldwide, yet diagnosis is often protracted and error-prone. Many conditions present with heterogeneous signs that overlap with common disorders, leading... Read more
AI-Driven Diagnostic Demonstrates High Accuracy in Detecting Periprosthetic Joint Infection
Periprosthetic joint infection (PJI) is a rare but serious complication affecting 1% to 2% of primary joint replacement surgeries. The condition occurs when bacteria or fungi infect tissues around an implanted... Read moreIndustry
view channel
Automated MSI Test Gains IVDR Certification to Guide CRC Therapy
Treatment selection for metastatic colorectal cancer often requires knowledge of a tumor’s microsatellite instability (MSI) status. Timely results can help clinicians decide on immunotherapy options.... Read more
