Eukaryote-Made Thermostable DNA Polymerase Enables Rapid PCR-Based Detection of Mycoplasma, Ureaplasma and Other Bacteria in the Amniotic Fluid of Preterm Labor Cases

摘要: Background Intra-amniotic infection has long been recognized as the leading cause of preterm delivery. Microbial culture is gold standard for detection intra-amniotic infection, but several days are required, and many bacterial species in amniotic fluid difficult to cultivate. Methods We developed a novel nested-PCR-based assay detecting Mycoplasma, Ureaplasma, other bacteria fungi samples within three hours sample collection. To detect prokaryotes, eukaryote-made thermostable DNA polymerase, which free from contamination, used combination with universal primers. In contrast, eukaryotes, conventional bacterially-made polymerase fungal assess validity PCR assay, we compared results using 300 samples. Results Based on level (positive negative), 93.3% (280/300) 94.3% (283/300) 89.3% (268/300) 99.7% (299/300) matched results. Meanwhile, concerning than Mycoplasma 228 were negative according method, 98.2% (224/228) also based method. Employing devised primer sets, mixed infections Ureaplasma and/or could be clearly distinguished. addition, attempted compare relative abundance 28 judged dominance by comparing Ct values quantitative real-time PCR. Conclusions rapid samples. This can applied accurately diagnose absence We believe that this will positively contribute treatment prevention