Molecular Identification of StaphylococcussppIsolatedfrom Clinical samples

Mohammad, TaghreedKhudhur

	Molecular Identification of StaphylococcussppIsolatedfrom Clinical samples
Jornal of Biotechnology Research Center
Article 1, Volume 12, Issue 1, 2018, Pages 14-24
Author
TaghreedKhudhur Mohammad
Abstract
The analysis of 16S rRNA gene sequences has been the technique generally used to study and confirm the identification andtaxonomy of staphylococci. However, bacterial species cannot always be distinguished from each other using cultural methods. Thus,clinicalsamples were collected from 190 cases only 31 positive for staphylococcal infections withUrinary Tract Infection, Wounds, Burns, Otitis media, diarrhea infections, were applied for microbiological analysis which include: cultures onManitol salt agar and HiCrome UTI Agar medium all the isolates gave positive golden yellowand identify as Staphylococcusspp. DNA was extracted fromStaphylococcussppand the 16srRNA gene were amplified by using specific primer, then sequencing of nucleic acid of genes was performed by machine is AB13730XL, Applied Biosystem, Macro gen company,the DNA sequencing results of flank sense of 16srRNA gene from 31strains of Staphylococcuswas confirm the identification into species level: Staphylococcus haemolyticus, Staphylococcus aureus, Staphylococcus epidermidisAnd Staphylococcus sciuri.Analysis of the sequences appeared that there two substitution(Transversion, Transition) in the Staphylococcusaureusstrains with Sequence IDLC090540.1 location at Range of nucleotide from 4 to 636, 100% compatibility with NCBI while no substitution appeared in the Staphylococcus haemolyticusstrains which have thesequence IDLN998078.1, 99% compatibility with NCBIalso the sequence ID KR812401.1 which related to the strain Staphylococcussciuri not appeared any substitution after sequencing analysis.Types of substitution detected in partial16srRNA gene inStaphylococcus epidermidisstrains 13 Transversion and 5 transition substitutionlocation at range of nucleotide from 6 to 1026 have the Sequence IDKF575160.1 compared with data obtained from Gene Bank, these finding lead to conclusion,our assay allows rapid and confirm the detection to avoid possibility of misidentification of Staphylococcus species based on cultural analysis, the study aimed to propose the partial sequencing of the gene as an alternative molecular tool for the analysis of Staphylococcus species and for decreasing thepossibility of misidentification. New submission of local Iraqi Staphylococcus clinical isolated during the current study show successfully record of four isolate Staphylococcus sciuri,Staphylococcus epidermidis,Staphylococcus aureus andStaphylococcus haemolyticuswith GenBank accession number: KY938530.1,KY938529.1,,KY938528.1, and GenBank: KY938527.1respectivelly.
Keywords
Key Words; HiCrome UTI Agar; Manitol salt Agar; and Sequencing; Gen Bank

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