The confirmation tests, carried out under optimum conditions, revealed that the experimental results were in harmony with the expected values from the Taguchi model.
The agglomerate yield (91.03%) obtained during the confirmation experiment carried out under optimum conditions was much higher than that observed in all the test runs and thereby, the authenticity of optimization was checked.
Further detection and confirmation was carried out according to CDC and WHO guidelines.
PCR confirmation was carried out using DNA extracted from healthy potatoes (negative control) and potatoes exhibiting symptoms of ZC disease.
RAD haplotype confirmation was carried out as above except that the forward primer was within the RAD site sequence.
Mice with CNS-specific deletions of Ptc1 were obtained by breeding animals carrying the conditional Ptc1 (Ptc1Lor/Lox) [22] oRbpjpj allele (RbpjLox/Lox) [24] with Nestin-Cre (NestinCre) transgenic mice [23] and genotyping confirmation was carried out as outlined.
For independent confirmation, we carried out immunoisolation of synaptic vesicles using beads (Eupergit C1Z) covalently coupled with monoclonal antibodies specific for Rab26 or synaptophysin.
Biopsies confirmation was carried out whenever possible and the proof of invasion of central nervous system (CNS) was ascertained by positive cytologic findings in cerebrospinal fluid.
In addition, when necessary, further confirmation was carried out with VITEK MS, a matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) system (bioMerieux, Marcy-l'Etoile, France).
After receipt by the central laboratory, International Health Management Associates, Inc. (IHMA, Schaumberg, IL, USA), organism identification confirmation was carried out on all isolates using matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (Bruker Daltronics, Bremen, Germany).
