Un método innovador basado en microplacas para medir la coagregación entre un par modelo de bacterias orales que se...

Coaggregation is proposed to enhance oral biofilm development. This study aims to evaluate an innovative high-throughput spectrophotometric method for quantifying coaggregation, which relies upon dome-shaped wells (DSWs) in microplates to partition coaggregates in cell suspensions, and compare its utility against other established methods.

Batch-cultured cells of Streptococcus gordonii DL1 and Actinomyces oris T14V were suspended individually in coaggregation buffer (Coag-B) or buffered KCl (B-KCl) for coaggregation experiments. Multiple methods were used to assess coaggregation between S. gordonii DL1 and A. oris T14V in the two buffers: the high-throughput spectrophotometric assay using modified 24-well microplates; visual aggregation assays; confocal microscopy; and cuvette-based spectrophotometry.

In either buffer, S. gordonii DL1 and A. oris T14V coaggregated strongly in visual tube-based assays, although the coaggregates were typically larger and more condensed in B-KCl. This observation was supported by microscopy and cuvette-based measurements. Compared with other well/buffer combinations, coaggregation was most strongly and reproducibly detected in endpoint and kinetic studies when cells were suspended in B-KCl and added to DSWs.

Using DSWs in combination with B-KCl provided a high-throughput quantitative approach to study coaggregation between S. gordonii DL1 and A. oris T14V.