Kidney stone disease is a common disease which affecting the population with a peak of incidence around the third to fourth decade of life. The present study deals with isolation, identification and analysis of bacteria from kidney stones through 16S rRNA based molecular technique. Bacterial strain was isolated and characterized using various biochemical tests and confirmed through molecular approach. Bacterial 16S rRNA gene was amplified using suitable primers. The amplified 16S rRNA gene sequence was compared with the sequence in NCBI sequence database the bacterial strain was identified as Acinetobacter calcoaceticus strain phylogenetic and molecular evolutionary analyses were conducted using 16S rRNA sequencing. The sequence when submitted to NCBI gene bank database using BLAST showed 99 – 100% maximum identity and has shown good similarity with Acinetobacter colcoaceticus, Acinetobacter oleivorans and Acinetobacter baumanni.
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