| Summary: | (1) Background: The wet mount staining technique is a simple, economical, and rapid method for detecting parasitic stages. The objective of the current study was to evaluate wet mount diagnostic stains for <i>Cryptosporidium</i> infection in human faecal samples and to compare the sensitivity and qualitative performance of toluidine blue (TolB) and modified Ziehl–Neelsen (mZN) stain. (2) Methods: The collection, purification, and molecular amplification of <i>Cryptosporidium</i> oocysts were performed. TolB, malachite green, trypan blue, and crystal violet were evaluated qualitatively to diagnose <i>Cryptosporidium</i> oocysts. The outperforming stain was compared to mZN using a quantitative evaluation and qualitative scoring system. (3) Results: The oocysts of <i>Cryptosporidium parvum</i> were effectively purified and utilized for spiking. TolB was the most effective diagnostic stain among wet mount stains for detecting <i>Cryptosporidium</i> oocysts. TolB outperformed mZN in terms of sensitivity, with 100% versus 79% at a concentration of 10<sup>4</sup> and 49% versus 23% at a concentration of 10<sup>2</sup>. TolB had the most favourable qualitative stain characteristics against mZN; however, sample freshness and being a temporary stain were crucial considerations. (4) Conclusions: This study emphasizes that TolB, as a routine wet mount technique for screening <i>Cryptosporidium</i> infection, will provide a more sensitive and faster method than mZN staining.
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