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Evaluation of real-time PCR based on SYBR Green I fluorescent dye for detection of Bacillus anthracis strains in biological samples
2018
Kędrak-Jabłońska, Agnieszka | Budniak, Sylwia | Szczawińska, Anna | Reksa, Monika | Krupa, Marek | Szulowski, Krzysztof
Introduction: The aim of the study was the application and evaluation of real-time PCRs based on the fluorescence of SYBR Green I intercalating dye for the detection of three Bacillus anthracis genes in contaminated liver and blood samples. The goals for detection were rpoB gene as a chromosomal marker, pag gene located on plasmid pXO1, and capC gene located on plasmid pXO2. Material and Methods: Five B. anthracis strains were used for the experiments. Additionally, single strains of other species of the genus Bacillus, i.e. B. cereus, B. brevis, B. subtilis, and B. megaterium, and strains of six other species were used for evaluation of the specificity of the tests. Three SYBR Green I real-time PCRs were conducted allowing confirmation of B. anthracis in the biological samples. Results: The observation of amplification curves in real-time PCRs enabled the detection of the chromosomally encoded rpoB gene, pag gene, and capC gene of B. anthracis. The specificity of the tests was confirmed by estimation of the melting temperature of the PCR products. The sensitivity and linearity of the reactions were determined using regression coefficients. Strains of other microbial species did not reveal real-time PCR products. Conclusion: All real-time PCRs for the detection of B. anthracis in biological samples demonstrated a significant sensitivity and high specificity.
Show more [+] Less [-]Fractionated RBD palm stearin as potential rumen bypass supplements
2012
Norliza Saparin | Zahariah Ismail | Mohd Suria Affandi Yusoff | Shawaluddin Tahiruddin
Refined, Bleached and Deodorized Palm Stearin (RBDPS) is a solid fraction obtained from Refined, Bleached and Deodorized Palm Oil after fractionation by crystallization at controlled temperature. Fractionated RBDPS was found to be enriched with C16 fatty acids,
lacking in trans fat and has a high melting point. The fractionated RBDPS produced had an iodine value of 13.1 gI2/100g, C16 content of 79.7% and melting point 60°C .These characteristics indicate fractionated RBDPS has potential as a rumen bypass fat as well as providing desirable carbon chain composition for good milk characteristics.
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