However, the frequency of conclusive samples using this method varies and is often too low, especially when performed by unexperienced operators.
In this study we have developed and tested a new semi-automated instrument ("CytoTest") designed for FNA which is intended to improve the efficacy of the technique by increasing the percentage of conclusive samples.
All these techniques were first set up on healthy and BSE non autolytic samples prior to be applied on autolytic control healthy and the five non-conclusive samples afterwards.
All the confirmatory techniques established by the OIE (immunohistochemistry, Western blotting, SAF visualization by electron microscopy and bioassay in a murine model) were applied on problem non-conclusive samples and were modified in order to increase the sensitivity to detect foreseeable low concentrations of PrPsc.
Derivation of the anchor-based thresholds of meaningful change might have been more conclusive if sample size was larger for the minimally-improved and minimally-worsened groups.
All embryologists performed similarly in terms of conclusive CCS diagnosis, sample concurrence and number of TE cells sampled from the different blastocysts.
We present a specific protocol which eliminates the need for time-consuming and expensive genomic DNA extractions and, using a high-processivity polymerase, demonstrate conclusive screening of samples in <1 h.
Therefore, results for dissolution testing were positive and conclusive for 123 samples.
Nevertheless, we recognize that this result is not conclusive because the sample size of the non-STG was relatively small.
For some of the analyzed samples, conclusive results were not obtained following the real-time PCR analysis, due to the presence of some inhibitory compounds in the DNA extracts (B. edulis, ascending segment; P. ostreatus, transverse and distal segments).
However, we need to underline that these are early results that need to be extended for more combinations, codebook sizes and types of collections, in order to draw conclusive statements about appropriate sampling rates.
