Each other strands used the same group of internal primers as with respective PCR assays
dos.4.3. Sequencing away from DNA
To have DNA sequencing, the fresh amplification facts obtained from this new PCR reactions were purified having the fresh Separate II PCR and you may Solution System (Bioline, London area, UK) and sequenced from inside the a keen Avant 3100 sequencer (Used Biosystem, Waltham, MA, USA). DNA sequencing are did from the a sequencing facility at the University off Veterinary Medication, Kosice, Slovakia. The new gotten DNA sequences from the partial bg and you may tpi family genes had been in contrast to new site sequences away from G. duodenalis regarding GenBank databases by nucleotide BLASTn program . Sequences was sorted from the resemblance and you may aimed utilising the Clustal Omega tool . Phylograms was indeed built from the MEGA7 software making use of the restrict-probability (ML) method having 1000 bootstrap pseudoreplicates. Good TN93 replacing model having gamma-marketed rate heterogeneity try chosen in line with the limit possibilities analysis of you’ll replacing habits making use of the Modeltest in Mega.
dos.5. Statistical Data
Statistical analysis of the obtained results was performed using the statistical product StatSoft, Inc. (2007). STATISTICA (software system for data analysis), version 8.0. and StatSoft Statistica 12 and Microsoft Excel (Office 365). The potential risk was calculated using the exchange rate (OR) as a relative chance and the relative risk using the risk rate (RR). Crosstables and Chi-square tests were used to calculate correlations between sex, the children’s residence belonging to a majority or minority population, and age-class. (далее…)