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Establishment of the molecular beacon-loop-mediated isothermal amplification method for the rapid detection of Porphyromonas gingivalis

Su, Yuxin, Huang, Simo, Hong, Lei, Zou, Dayang, Tang, Yue, Chao, Siqi, He, Xiaoming, Xu, Yaqing, Liu, Xinwei, Li, Lun, Feng, Lili, Li, Wenfeng, Liu, Wei, Ke, Yuehua, Huang, Liuyu
Journal of microbiological methods 2019 v.160 pp. 68-72
Porphyromonas gingivalis, cross reaction, diagnostic sensitivity, pathogens, periodontal diseases, point-of-care systems, quantitative polymerase chain reaction, rapid methods
Porphyromonas gingivalis, a clinically important oral pathogen causing periodontal disease, is difficult to culture in routine conditions. Hence, it is necessary to establish a reliable technique to detect this pathogen. Previously, our laboratory developed a new isothermal detection method, called MB-LAMP (molecular beacon-Loop-mediated isothermal amplification), which combines the advantages of LAMP and qPCR through the accurate and quantitative detection of LAMP products. This approach offers significant potential for the point-of-care detection of P. gingivalis. Here, MB-LAMP was used to detect P. gingivalis targeting a specific fragment, and the sensitivity was as high as 1.4 × 10−1 pg μL–1. The method showed no cross-reaction with 14 other bacterial pathogens. For clinical samples, this assay showed a high diagnostic sensitivity (100%) and specificity (100%), equivalent to that of real-time quantitative polymerase chain reaction (real-time qPCR). Moreover, detection with MB-LAMP was significantly faster than that with real-time qPCR, reducing the time required for clinical diagnosis. Finally, we established an absolute quantification method with MB-LAMP for P. gingivalis using pilot samples. Thus, the highly specific, sensitive, and rapid assay developed in this study makes it feasible to diagnose P. gingivalis.