AI Article Synopsis
- * The microscopic analysis involved staining with vital dyes, while the molecular analysis assessed DNA amplification after applying PMA to the oocysts.
- * Results indicate that qPCR-PMA is an effective technique for identifying whether T. gondii oocysts in water are viable or non-viable, which is essential for determining water safety.
Article Abstract
This study aimed to investigate a methodology for discriminating viable and non-viable T. gondii oocysts in water. Analyses included two steps: (i) microscopic investigation with vital dyes; (ii) molecular investigation, using a real time PCR (qPCR), after parasite treatment (or not) with propidium monoazide (PMA). The method was called qPCR-PMA. Oocyst aliquots were incubated (15 min) at 25 ºC or 100 ºC and analyzed by microscopy, after trypan blue and neutral red staining. Microscopic investigation determined viable and non-viable oocysts. For the molecular investigation, both aliquots of oocysts were treated with PMA. Non-viable oocysts, after PMA treatment, exhibited an inhibition of DNA amplification by qPCR. Although analyses were carried out with oocysts treated experimentally, these results suggest that qPCR-PMA can be a useful strategy to distinguish viable and non-viable T. gondii oocysts in water safety testing, showing if water is safe to drink.
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|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7608075 | PMC |
| http://dx.doi.org/10.1590/S1678-9946202062084 | DOI Listing |
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