Microsporidia PCR detection artifacts due to non-specific binding of the universal microsporidia primers to the rDNA of arthropod hosts
2010
Tokarev, Yu., All-Russia Research and Development Inst. of Plant Protection, Saint-Petersburg (Russian Federation) | Sitnikova, N. | Pistone, D.
For amplification and sequencing of the microsporidian rDNA, "universal microsporidia primers" have been designed and efficiently exploited in a number of works to identify dozens of species of these animal pathogens. The data presented in this paper clearly demonstrate that the primer 18f:530r and 18f:1492r pairs used in PCR under conditions described may result in amplification of the host, not microsporidian rDNA. The efficacy of this amplification is far from 100 percent (despite the fact that the arthropod rDNA must be abundantly present in all the DNA samples screened as it was isolated from the arthropod tissues), thus providing false positives at the prevalence levels typical for microsporidian infections in the natural populations of arthropods. It is obvious that the reverse primers are annealed in a way that is not characteristic of the arthropod rDNA though it is not clear what conditions ensure annealing of the forward primers that are not homologous to the respective arthropod rDNA
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