Quantitative PCR for Etiologic Diagnosis of Methicillin-Resistant Staphylococcus aureus Pneumonia in Intensive Care Unit

Tuberculosis and Respiratory Diseases

article

Sun-Jung Kwon¹  Taehyeon Jeon¹  Dongwook Seo¹  Moonjoon Na²  Eu-Gene Choi²  Ji-Woong Son²  Eun-Hyung Yoo³  Chang-Gyo Park¹  Hoi Young Lee¹  Ju Ock Kim⁴  Sun-Young Kim⁴  Jaeku Kang¹ 

[1] Myunggok Medical Research Institute, Konyang University College of Medicine;Department of Internal Medicine, Konyang University Hospital, Konyang University College of Medicine;Department of Laboratory Medicine, Konyang University Hospital, Konyang University College of Medicine;Department of Internal Medicine, Chungnam National University School of Medicine

关键词: Methicillin-Resistant Staphylococcus aureus;    Real-Time Polymerase Chain Reaction;    Pneumonia;    Ventilator-Associated;   

DOI  :  10.4046/trd.2012.72.3.293

学科分类：医学（综合）

来源: The Korean Academy of Tuberculosis and Respiratory Diseases

【 摘 要 】

Background Ventilator-associated pneumonia (VAP) requires prompt and appropriate treatment. Since methicillin-resistant Staphylococcus aureus (MRSA) is a frequent pathogen in VAP, rapid identification of it, is pivotal. Our aim was to evaluate the utility of quantitative polymerase chain reaction (qPCR) as a useful method for etiologic diagnoses of MRSA pneumonia. Methods We performed qPCR for mec A, S. aureus -specific fem A-SA, and S. epidermidis -specific fem A-SE genes from bronchoalveolar lavage or bronchial washing samples obtained from clinically-suspected VAP. Molecular identification of MRSA was based on the presence of the mec A and fem A-SA gene, with the absence of the fem A-SE gene. To compensate for the experimental and clinical conditions, we spiked an internal control in the course of DNA extraction. We estimated number of colony-forming units per mL (CFU/mL) of MRSA samples through a standard curve of a serially-diluted reference MRSA strain. We compared the threshold cycle (Ct) value with the microbiologic results of MRSA. Results We obtained the mec A gene standard curve, which showed the detection limit of the mec A gene to be 100 fg, which corresponds to a copy number of 30. We chose cut-off Ct values of 27.94 (equivalent to 1×10 4 CFU/mL) and 21.78 (equivalent to 1×10 5 CFU/mL). The sensitivity and specificity of our assay were 88.9% and 88.9% respectively, when compared with quantitative cultures. Conclusion Our results were valuable for diagnosing and identifying pathogens involved in VAP. We believe our modified qPCR is an appropriate tool for the rapid diagnosis of clinical pathogens regarding patients in the intensive care unit.

【 授权许可】

CC BY-NC