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Molecular detection of virulance nuc gene of staphylococcus species isolated from sinusitis’s patients by multiplex PCR

Author: 
Maryam N. Hamad and Saife D. Al-Ahmer
Subject Area: 
Physical Sciences and Engineering
Abstract: 

The staphylococci are most frequently isolated from clinical specimens in the microbiology laboratory. These bacteria are widespread in nature and can be recovered from environment or as commensally inhabitants of the skin, mucous membranes and other body sites in humans and animals. Aim of the current study was to Identify the different coagulase positive (CoPS) and coagulase negative (CoNS) Staphylococcus species includes S. aureus, S. lugdunensis, S. epidermidis, and S. hominis isolated from sinusitis’s patients by multiplex PCR. In this study nasal swabs from 150 patients were used for culture, VITEK 2 system and multiplex PCR analysis. Multiplex PCR was performed with species-specific primers targeted to the nuc gene of S. aureus, S. lugdunensis, S. epidermidis, and S. hominis. The result of the multiplex PCR was compared with conventional culture,VITEK 2 system methods. The positive multiplex PCR product was identified by presence of ~359 bp, ~659 bp, ~251 bp and ~ 177 amplicons of nuc gene for the S. aureus, S. lugdunensis, S. epidermidis, and S. hominis, reaspectively. Conventional methods of Staphylococcus culture, VITEK 2 system, showed that to sum up 100 out of 150 nasal swabs were detected for Staphylococcus species; To sum up 31 out of 100 nasal swabs were detected for S. aureus, 16 out of 100 nasal swabs were detected for S. lugdunensis, 10 out of 100 nasal swabs were detected for S. epidermidis, 6 out of 100 nasal swabs were detected for S. hominis and 37 out of 100 nasal swabs were detected for other Staphylococcus species. Five of the 60 samples that were negative by conventional methods were positive by multiplex PCR. Statistical analysis revealed that the PCR to have a sensitivity of 95.5 % in the detection of Staphylococcus species in sinusitis. This multiplex PCR method provides a sensitive, rapid, and reliable alternative to conventional methods to identify Staphylococcus species isolated from patients clinically diagnosed with sinusitis.

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