TITLE:
Development of a Duplex Real-Time PCR Method for the Pharmaceutical Rapid Microbial Detection of Staphylococcus aureus and Pseudomonas aeruginosa
AUTHORS:
Tiehao Lin, Liying Lin, Pu Zeng
KEYWORDS:
S. aureus, P. aeruginosa, Duplex Real-Time PCR, Pharmaceutical
JOURNAL NAME:
Journal of Biosciences and Medicines,
Vol.2 No.5,
July
22,
2014
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.