Alfa M J, DeGagne P
St. Boniface Research Centre, Winnipeg, Manitoba, Canada.
Microb Pathog. 1994 Sep;17(3):167-74. doi: 10.1006/mpat.1994.1063.
This study evaluated a commercially available chemiluminescent-labelled, ribosomal RNA-directed DNA probe (CRP) as a method to quantitate attachment of H. ducreyi to human foreskin fibroblast (HFF) cells. Evaluation of four strains of H. ducreyi demonstrated that the CRP assay was unaffected by eukaryotic cells and its advantages were: (i) quantitation was done after attachment so it did not interfere with the attachment process, and (ii) it was a rapid, reliable method for quantitating bound bacteria, despite bacterial clumping. Gentamicin-killed H. ducreyi attached to both HFF cells and viable controls, suggesting that the adhesins are components constitutively present on the surface of H. ducreyi. This method may be widely applicable, since the probe recognizes most prokaryotic rRNA sequences.
本研究评估了一种市售的化学发光标记、核糖体RNA导向的DNA探针(CRP),作为定量检测杜克雷嗜血杆菌与人包皮成纤维细胞(HFF)附着的方法。对四株杜克雷嗜血杆菌的评估表明,CRP检测不受真核细胞影响,其优点如下:(i)在附着后进行定量,因此不干扰附着过程;(ii)尽管存在细菌聚集现象,但它是一种快速、可靠的定量结合细菌的方法。庆大霉素杀死的杜克雷嗜血杆菌附着于HFF细胞和活细菌对照,这表明黏附素是杜克雷嗜血杆菌表面组成性存在的成分。由于该探针可识别大多数原核rRNA序列,此方法可能具有广泛的适用性。