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Pathogens 2015, 4(2), 199-209; doi:10.3390/pathogens4020199

Direct Detection of Methicillin-Resistant Staphylococcus Aureus in Sputum Specimens from Patients with Hospital-Associated Pneumonia Using a Novel Multilocus Pcr Assay

1
State Key Laboratory of Respiratory Diseases, the first affiliated hospital of Guangzhou Medical University, Guangzhou, 510120, China
2
Peking University Health Science Center, Beijing, 100191, China
*
Author to whom correspondence should be addressed.
Academic Editor: Lawrence S. Young
Received: 26 February 2015 / Revised: 23 April 2015 / Accepted: 28 April 2015 / Published: 30 April 2015
View Full-Text   |   Download PDF [230 KB, uploaded 30 April 2015]

Abstract

Methicillin-resistant Staphylococcus aureus (MRSA) is a significant cause of hospital-associated pneumonia (HAP). The rapid identification of MRSA would be beneficial for early diagnosis. The study aimed to evaluate a multilocus, fluorescence-based PCR assay based on the detection of mecA and nuc genes for identification of S. aureusin lower respiratory tract (LRT) specimens. Sensitivity and specificity of the PCR assay were analyzed. Clinical evaluation for the assay was performed using LRT specimens from patients with HAP, and the sensitivity, specificity, positive and negative predictive values (PPV and NPV) were evaluated in comparison with semi-quantitative culture methods. The result showed the assay provided positive identification of all MRSA reference strains with a limit of detection for MRSA of 4 × 103 CFU/mL. Compared with semi-quantitative culture, the sensitivity, specificity, PPV and NPV were 100%, 89.6%, 75.0%, and 100%, respectively. A positive correlation between MRSA bacterial colonies and PCR copy number was found. The specificity and PPV reached 96.6% and 89.7% respectively, if the PCR copy number reached a definite positive threshold of 5.96 × 105. It suggested that this novel multilocus, fluorescence-based PCR assay proved to be a fast, sensitive and specific tool for direct detection of MRSA from LRT specimens. View Full-Text
Keywords: MRSA; mecA gene; nuc gene MRSA; mecA gene; nuc gene
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Huang, Z.-G.; Zheng, X.-Z.; Guan, J.; Xiao, S.-N.; Zhuo, C. Direct Detection of Methicillin-Resistant Staphylococcus Aureus in Sputum Specimens from Patients with Hospital-Associated Pneumonia Using a Novel Multilocus Pcr Assay. Pathogens 2015, 4, 199-209.

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