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
Platinum Member
Automated Coagulation Analyzer
Hemolumi H6
Gold Member
Nucleic Acid Extractor System
NEOS-96 XT
New
Gold Member
Blood-Based Protein Biomarker Solution for Alzheimer's Disease
BG-DTi2000.
Japanese Encephalitis Test
Japanese Encephalitis Virus Real Time PCR Kit
Latest Immunology News
- AI-Based Antibody Profiling Predicts Strength of COVID-19 Vaccine Response
- Immune Biomarkers May Predict Recurrent Checkpoint Inhibitor Arthritis
- Immune Cell Blood Test May Predict Melanoma Immunotherapy Response
- Study Reveals Viral Protein Driving COVID-19-Related Vascular Injury
- Antibody Profiling Identifies Preclinical Inflammatory Bowel Disease Years Before Diagnosis
- Study Reveals Immune Mechanism Driving Severe COVID-19 Progression
- Ultrasensitive Blood Test Detects Sjögren’s Signature Years Before Diagnosis
- New Assays Expand Cytokine Testing for Transplant and Immunocompromised Patients
- Cell-Free Assay Detects Functional IgE for Food Allergy Diagnosis
- Diagnostic Models Detect Hidden Eye Abnormalities After Mild COVID-19
- Anti-Lipid Antibody Biomarkers May Identify Early Lyme Disease and Persistent Symptoms
- Immune Biomarkers Could Identify Risk of Chronic Critical Illness on ICU Admission
- Emergency Department Opt-Out Testing Program Identifies Undiagnosed HIV
- Airway Immune Signature May Predict Tuberculosis Progression Risk
- New Cellular Biomarkers Correlate with Disease Severity in Sjögren Disease
- Lung Immune Profiling Reveals Distinct Severe Pneumonia Subtypes
Channels
Clinical Chemistry
view channel
Blood Test Markers Could Help Identify Older Adults at Risk of Disability
Maintaining independence into very old age is a growing public health challenge as Japan’s population rapidly ages. Nearly 60% of Japanese adults aged 85 years and older already receive support through... Read more
FDA Clears Roche Blood Test for Alzheimer’s Amyloid Pathology Assessment
An estimated 75% of people living with dementia remain undiagnosed, while confirmatory tools such as positron emission tomography (PET) or cerebrospinal fluid (CSF) testing can be costly, invasive, and... Read more
Rapid Ferritin Test Enables Iron Deficiency Detection at the Point of Care
Iron deficiency is one of the world’s most common nutritional disorders and can be difficult to diagnose where access to laboratories and trained personnel is limited. Women and children in low-resource... Read more
Alzheimer’s Blood Test Becomes First FDA-Cleared Option for Adults as Young as 40
C2N Diagnostics’ PrecivityAD2 blood test was cleared by the U.S. Food and Drug Administration (FDA) on August 20, 2026, for adults as young as 40 who are experiencing signs of cognitive impairment, according... Read moreMolecular Diagnostics
view channel
Genetic Risk Score Identifies Type 1 Diabetes in MODY Testing
Accurately distinguishing maturity-onset diabetes of the young from type 1 diabetes is difficult in routine care, especially when onset occurs in adolescence or early adulthood. Although maturity-onset... Read more
Self-Collected HPV Testing Reaches Patients Missed by Routine Screening
Timely cervical cancer screening prevents progression from persistent high‑risk human papillomavirus (HPV) infection, yet many adults marginalized from care do not complete routine exams.... Read more
Comprehensive Genomic Testing Expands Treatment Options for Advanced Cancers
Selecting effective therapies for advanced cancers remains difficult because standard tumor testing often targets only a limited set of genes. As options dwindle, many patients face decisions without strong... Read more
Multi-Omics Analysis Identifies Additional Risk Genes in Hereditary Breast and Ovarian Cancer
Hereditary breast and ovarian cancer can be difficult to explain genetically, leaving many high-risk families without clear answers. Although 13 established risk genes, including BRCA1 and BRCA2, are routinely... Read moreHematology
view channel
New Genetic Findings Reveal Cause of Bone Marrow Failure Syndrome
Inherited bone marrow failure syndromes (IBMFS) impair the bone marrow’s ability to produce sufficient healthy blood cells and are associated with an increased risk of early-onset myelodysplastic syndromes (MDS).... Read more
Ultra-Portable Device Enables Finger-Prick Blood Testing at Home
Patients who need frequent blood tests often face repeated clinic visits that burden services and disrupt care. In the UK, millions of tests are performed each year to diagnose disease, guide therapy,... Read more
Immunology
view channel
AI-Based Antibody Profiling Predicts Strength of COVID-19 Vaccine Response
Vaccine-induced protection can vary widely between individuals, creating uncertainty for clinicians, particularly in patients with immunosuppressive conditions. Demographic and health factors explain only... Read more
Immune Biomarkers May Predict Recurrent Checkpoint Inhibitor Arthritis
Inflammatory arthritis is a recognized immune-related adverse event in patients treated with immune checkpoint inhibitors (ICIs) for cancer. Between 20% and 50% of affected patients experience more than... Read more
Pathology
view channel
AI-Pathologist Framework Improves Accuracy and Reliability in Cancer Diagnosis
Ensuring reliable cancer diagnosis from digital pathology remains a critical challenge as clinical decisions rely on accurate slide interpretation. While artificial intelligence (AI) has accelerated whole-slide... Read more
New Review Highlights Intelligent Agents as Next Step for Digital Pathology
Digital pathology remains constrained by models that classify single images without mirroring how clinicians interrogate multiple slides, adjust magnification, and synthesize ancillary tests.... Read more
Simple Immunohistochemical Size Ratio Improves Classification of Primary Aldosteronism
Primary aldosteronism is a common but underdiagnosed cause of hypertension, in which excess aldosterone promotes salt retention and raises blood pressure. Identifying the dominant source of hormone overproduction... Read more
Technology
view channel
Autonomous Robotic System Receives FDA Authorization for Blood Collection
Venipuncture is central to many diagnostic pathways, but routine blood collection can be affected by staffing constraints and procedural variability that influence consistency and patient experience.... Read more
Interoperable Data Platform Standardizes Multi-Cancer Blood Test Results
Proteotype Diagnostics has introduced Alchemi, a proprietary data and clinical workflow platform being developed for Enlighten, the company’s investigational blood-based multi-cancer test.... Read more
Industry
view channel
New Collaboration Advances Precision Oncology Screening for Lung Cancer in Japan
Molecular profiling is central to precision oncology in lung cancer, but tissue samples can be limited and minimally invasive approaches are often preferable. Liquid biopsy enables blood-based genomic... Read more
