Peptoniphilus mikwangii-specific quantitative real-time polymerase chain reaction primers

Soon‐Nang Park, J. Kook
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Abstract

The purpose of this study was to develop Peptoniphilus mikwangii -specific quantitative real-time polymerase chain reaction (qPCR) primers based on the 16S ribosomal RNA (16S rDNA) gene. The specificity of the primers was determined by conventional PCR using 29 strains of 27 oral bacterial species including P. mikwangii. The sensitivity of the primers was determined by qPCR using the purified genomic DNA of P. mikwangii KCOM 1628T (40 ng to 4 fg). The data showed that the qPCR primers (RTB134-F4/RTB134-R4) could detect P. mikwangii strains exclusively and as little as 40 fg of the genomic DNA of P. mikwangii KCOM 1628T. These results suggest that the developed qPCR primer pair can be useful for detecting P. mikwangii in epidemiological studies of oral bacterial infectious diseases.
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米康氏胃杆菌特异性定量实时聚合酶链反应引物
本研究旨在构建基于16S核糖体RNA (16S rDNA)基因的米旺氏胃杆菌特异性定量实时聚合酶链反应(qPCR)引物。引物的特异性采用常规PCR方法对包括米旺氏假单胞菌在内的27种口腔细菌29株进行检测。用纯化的mikwangii KCOM 1628T基因组DNA (40 ng ~ 4 fg)进行qPCR检测引物的敏感性。结果表明,qPCR引物(RTB134-F4/RTB134-R4)能特异检测米王假单胞菌KCOM 1628T的基因组DNA,且只检测到40 fg。上述结果提示,所构建的qPCR引物对可用于口腔细菌性传染病的流行病学研究。
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