Yi Jongyoun, Yoon Bo Hyun, Kim Eui-Chong
Department of Laboratory Medicine, Seoul National University College of Medicine, 28 Yongon-dong, Chongnogu, Seoul 110-744, South Korea.
Mol Cell Probes. 2005 Aug;19(4):255-60. doi: 10.1016/j.mcp.2005.04.002.
Prenatal intrauterine infection has been recognized as an important cause of premature birth, and Ureaplasma urealyticum is one of the commonest pathogens. U. urealyticum consists of 14 serovars that can be divided into two biovars (parvo and T960), and the pathogenicity of U. urealyticum may be different according to the biovar. To detect U. urealyticum and determine its biovar simultaneously, we developed a real-time polymerase chain reaction (PCR) assay targeting urease gene. The real-time PCR biovar-typed two reference strains and 42 culture isolates of U. urealyticum as correctly as conventional PCR with direct sequencing. Subsequently, 87 clinical specimens (amniotic fluid, cord blood, vaginal swab) were tested for culture, conventional PCR, and real-time PCR. When compared with conventional PCR, sensitivity and specificity of real-time PCR were 89.5 and 98.5%, respectively, and those of culture were 47.4 and 100%, respectively. Of 18 clinical specimens that were found positive and biovar-typed by real-time PCR, parvo biovar was 66.7% and T960 biovar was 33.3%. This real-time PCR assay can be useful for the simultaneous detection and biovar discrimination of U. urealyticum in clinical specimens. Further study to quantify U. urealyticum would be facilitated on the basis of this method.
产前宫内感染已被公认为早产的重要原因，解脲脲原体是最常见的病原体之一。解脲脲原体由14个血清型组成，可分为两个生物型（细小生物型和T960生物型），解脲脲原体的致病性可能因生物型而异。为了同时检测解脲脲原体并确定其生物型，我们开发了一种针对脲酶基因的实时聚合酶链反应（PCR）检测方法。实时PCR对两株参考菌株和42株解脲脲原体培养分离株进行生物型分型的结果与传统PCR直接测序法一样准确。随后，对87份临床标本（羊水、脐血、阴道拭子）进行了培养、传统PCR和实时PCR检测。与传统PCR相比，实时PCR的敏感性和特异性分别为89.5%和98.5%，培养法的敏感性和特异性分别为47.4%和100%。在实时PCR检测呈阳性并进行生物型分型的18份临床标本中，细小生物型占66.7%，T960生物型占33.3%。这种实时PCR检测方法可用于临床标本中解脲脲原体的同时检测和生物型鉴别。基于该方法将有助于进一步开展解脲脲原体定量研究。
