Journal of Biosciences and Medicines

Volume 2, Issue 5 (July 2014)

ISSN Print: 2327-5081   ISSN Online: 2327-509X

Google-based Impact Factor: 0.87  Citations  

Development of a Duplex Real-Time PCR Method for the Pharmaceutical Rapid Microbial Detection of Staphylococcus aureus and Pseudomonas aeruginosa

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DOI: 10.4236/jbm.2014.25002    2,902 Downloads   3,971 Views  Citations

ABSTRACT

Objective: To develop a duplex real-time PCR assay for pharmaceutical rapid microbial detection of Staphylococcus aureus and Pseudomonas aeruginosa. Methods: The specific primers and probes were designed to amplify the femB gene of S. aureus and the DNA gyrase subunit B gene of P. aeruginosa. The sensitivity of the system was detected by a multiple proportional dilution method. In order to examine the specificity of the system, other twenty-one bacteria strains were assayed simultaneously. Results: A highly sensitive and specific duplex real-time PCR assay for the detection of S. aureus and P. aeruginosa was established. The sensitivity was 50 copies/μL. The specificity was 100%. The whole detection procedure can be finished within 2.5 h. Conclusion: The duplex real-time PCR method is efficient in detecting with good sensitivity and specificity. There is a good prospect of this method applying in disease prevention and pharmaceutical industry due to the simultaneous detection of two pathogens.

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Lin, T. , Lin, L. and Zeng, P. (2014) Development of a Duplex Real-Time PCR Method for the Pharmaceutical Rapid Microbial Detection of Staphylococcus aureus and Pseudomonas aeruginosa. Journal of Biosciences and Medicines, 2, 12-19. doi: 10.4236/jbm.2014.25002.

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