The goal of this project was to develop a rapid molecular technique that can specifically
detect and genotype viable Cryptosporidium oocysts. The authors have developed and optimized
the use of propidium monoazide (PMA) with a PCR-based assay that enables detection and
genotyping of viable oocysts. PMA is a DNA intercalating dye that only penetrates dead or
damaged cells. Unlike other intercalating "vital" dyes, PMA can covalently bind to DNA upon
exposure to bright light. As a result, only the genomic DNA from viable cells, which is not
intercalated with PMA, is retained following DNA extraction procedures and can subsequently
be detected by PCR analysis. To date, PMA has only been used to differentiate live/dead
bacteria and fungi and has not yet been applied to protozoan parasites. Results in this
paper show that heat killed oocysts treated with PMA were not detected while live oocysts
treated with PMA were detected using conventional PCR. Includes 7 references.
| Edition : | Vol. - No. |
| File Size : | 1
file
, 700 KB |
| Note : | This product is unavailable in Ukraine, Russia, Belarus |
| Number of Pages : | 3 |
| Published : | 11/01/2008 |
