The sensitivity and the small sample requirements make the assay suitable for high-throughput screening of biological samples.
In order to make the assay more accessible to a resource-limited clinical setting, we eliminated the ELAST system, which simplified the Up24 assay, reduced its cost, and tested the accuracy of the modified assay in a rural Malawian hospital.
The next step is to make the assay reproducibly quantitative.
To make the assay robust for clinical use a chemical compound was applied to the samples that inhibited interference from the classical pathway due to the presence of anti-BSA antibodies in some sera.
Further refinements could make the assay suitable for high-throughput screening.
Nine OSNA +I patients were excluded, owing to the presence of inhibiting materials, which make the assay less reliable.
Note that we have also added the PCR primers used for genotyping to the diagram to make the assay in Figure 1D clearer.
In order to make the assay as efficient as possible, the shortest possible AB reduction endpoint was determined as for S. epidermidis [ 14].
Because this approach will not require labor-intensive sand fly rearing and salivary gland dissection, this modification will make the assay more amenable for large-scale application.
To make the assay as rapid as possible, we evaluated phenotypes using primary colonies derived from independently transformed protoplasts and selected for stable antibiotic resistance.
