Validated cost-effective casein-based blocking buffers for enhanced neurocysticercosis ELISA accuracy

Area of Science:

• * Parasitology and Infectious Diseases
• * Immunodiagnostics and Assay Development

Background:

• * Neurocysticercosis (NCC) diagnosis requires accurate and affordable tools, especially in resource-limited settings.
• * Blocking buffers are critical in indirect ELISA (iELISA) to minimize background noise and ensure test reliability.
• * Limited comparative data exists on blocking buffer performance specifically for NCC iELISA.

Purpose of the Study:

• * To evaluate and compare the performance of various commercial and in-lab prepared blocking buffers for NCC indirect ELISA.
• * To identify cost-effective blocking solutions without compromising diagnostic accuracy.

Main Methods:

• * Nine blocking solutions (4 commercial, 5 in-lab) were tested using indirect ELISA.
• * Crude Cysticercus cellulosae antigen and a panel of 30 human serum samples (14 positive, 16 negative) were utilized.
• * All assay variables, except the blocking buffer, were kept constant to isolate buffer performance.

Main Results:

• * Six blocking formulations achieved 100% sensitivity and specificity.
• * Remaining buffers showed sensitivities between 84.6% and 93.7%.
• * A 3% casein-based in-lab buffer (B9) demonstrated excellent analytical consistency and diagnostic accuracy (AUC 1.000), with over 90% cost reduction.

Conclusions:

• * Well-prepared in-lab blocking buffers, particularly casein-based ones, can achieve diagnostic performance comparable to or exceeding commercial options in NCC iELISA.
• * This highlights a viable strategy for developing highly accurate and significantly more affordable diagnostic tools for neurocysticercosis.
• * The findings support the potential for improved access to NCC diagnostics in resource-limited settings.