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

DOI: 10.4236/jbm.2014.25002   PDF   HTML   XML   2,899 Downloads   3,965 Views   Citations


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.

Conflicts of Interest

The authors declare no conflicts of interest.


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