Advanced Detection Method for Dengue NS1 Protein Using Ultrasensitive ELISA with Thio-NAD Cycling.

Area of Science:

• Medical Diagnostics
• Virology
• Biochemistry

Background:

• Dengue virus (DENV) infection poses a significant global health and economic challenge.
• Current diagnostic methods for DENV are often slow, costly, or require specialized expertise, hindering early detection.
• The dengue virus non-structural protein 1 (NS1) is an early and reliable biomarker for DENV infection.

Purpose of the Study:

• To develop and evaluate an ultrasensitive enzyme-linked immunosorbent assay (ELISA) for detecting the dengue virus non-structural protein 1 (NS1).
• To assess the diagnostic performance of the developed ultrasensitive ELISA for dengue fever using clinical patient samples.

Main Methods:

• Development of a thio-NAD cycling ELISA for enhanced sensitivity in NS1 protein detection.
• Validation of the ultrasensitive ELISA using 85 clinical specimens (60 DENV-positive, 25 DENV-negative) compared against Nucleic Acid Amplification Test (NAAT) confirmation.

Main Results:

• The ultrasensitive ELISA achieved a limit of detection (LOD) of 1.152 pg/mL for NS1 protein.
• The assay demonstrated high clinical performance with 98.3% sensitivity and 100% specificity.
• Results showed excellent agreement with NAAT, indicated by a kappa coefficient of 0.972.

Conclusions:

• The ultrasensitive ELISA is a highly sensitive and specific method for detecting dengue virus NS1 protein.
• This assay presents a low-cost, user-friendly alternative to existing diagnostic methods, suitable for early dengue fever detection.
• The developed diagnostic tool holds significant potential for improving dengue management, particularly in low-income countries.