用于快速检测β-内酰胺酶阴性、氨苄西林耐药流感嗜血杆菌的多重实时聚合酶链反应

Multiplex real-time polymerase chain reaction for rapid detection of beta-lactamase-negative, ampicillin-resistant Haemophilus influenzae.

作者信息

Nakamura Shigeki, Yanagihara Katsunori, Morinaga Yoshitomo, Izumikawa Koichi, Seki Masafumi, Kakeya Hiroshi, Yamamoto Yoshihiro, Kamihira Shimeru, Kohno Shigeru

机构信息

Second Department of Internal Medicine, Nagasaki University Graduate School of Medical Science, 852-8501 Nagasaki, Japan.

出版信息

Diagn Microbiol Infect Dis. 2009 May;64(1):64-9. doi: 10.1016/j.diagmicrobio.2009.01.023. Epub 2009 Feb 20.

DOI:10.1016/j.diagmicrobio.2009.01.023

PMID:19232868

Abstract

We evaluated a multiplex real-time quantitative polymerase chain reaction (PCR) method for quantification of Haemophilus influenzae and rapid detection of beta-lactam-resistant strains. We designed 5 PCR primer sets to simultaneously detect the beta-lactam-resistant genes and quantify the pathogen. To demonstrate the validity of this assay, we used 191 clinical isolates, including 141 H. influenzae strains, and 100 purulent sputum samples, including 30 samples from which H. influenzae had been isolated. This assay showed 92.9% sensitivity and 91.8% specificity for detecting beta-lactam-resistant genes, relative to the conventional phenotypic method, and this assay correlated well with conventional quantitative culture counts. By using this assay, we could quantify H. influenzae and identify beta-lactam susceptibility in only 3 h and with only one tube. This method will be helpful for the rapid detection of H. influenzae infections and the selection of appropriate antibiotics.

摘要

我们评估了一种用于流感嗜血杆菌定量及β-内酰胺耐药菌株快速检测的多重实时定量聚合酶链反应（PCR）方法。我们设计了5组PCR引物，以同时检测β-内酰胺耐药基因并对病原体进行定量。为证明该检测方法的有效性，我们使用了191株临床分离株，其中包括141株流感嗜血杆菌菌株，以及100份脓性痰样本，其中包括30份已分离出流感嗜血杆菌的样本。相对于传统表型方法，该检测方法检测β-内酰胺耐药基因的灵敏度为92.9%，特异性为91.8%，且该检测方法与传统定量培养计数结果相关性良好。通过使用该检测方法，我们仅需3小时且仅用一管即可对流感嗜血杆菌进行定量并鉴定β-内酰胺敏感性。该方法将有助于快速检测流感嗜血杆菌感染并选择合适的抗生素。