文献类型：期刊文献

英文题名：Precise and simultaneous enumeration of multiplex pathogens using multiplex polymer chain reaction coupled with a novel quantitative capillary electrophoresis

作者：Liu, Chenchen[1];Yamaguchi, Yoshinori[1,2];Sekine, Shinichi[3];Ni, Yi[1];Dou, Xiaoming[1,2]

机构：[1]ECUST, Grad Sch Sci, Inst Photon & Biomed IPBM, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Osaka Univ, Grad Sch Engn, Dept Appl Phys, Suita, Osaka 5650871, Japan;[3]Osaka Univ, Grad Sch Dent, Dept Prevent Dent, 1-8 Yamadaoka, Suita, Osaka 5650871, Japan

年份：2018

卷号：258

起止页码：263

外文期刊名：SENSORS AND ACTUATORS B-CHEMICAL

收录：;EI(收录号：20174804458047);WOS:【SCI-EXPANDED(收录号:WOS:000424868700031)】；

基金：This project was partly supported by Grand-in-Aid for Scientific Research (A15H038270(Y.Y)), JSPS, japan and National Natural Science Foundation of China(No. 21305089). We acknowledged on partly financial support by East China University of Science and Technology (No. YK0142119).

语种：英文

外文关键词：Cell counting; Pathogen diagnosis; Quantitative capillary electrophoresis; Periodontal diseasea

摘要：High throughput, reliable and low cost methodologies for parallel pathogen counting are demanded in the diagnosis of infections, but still technically challenging. This work proposed a precise, simultaneous process of enumerating multiplex periodontal pathogens by using multiplex polymer chain reaction( mPCR) and quantitative capillary electrophoresis (qCE). A self-developed qCE protocol, which was time modified internal standard (TMIS) method, was employed to quantitatively study mPCR reactions. The optimal mPCR conditions were obtained for the amplification of three important periodontal pathogens, Porphyromonas gingivalis (P.g.), Treponema denticola (T.d.) and Tannerella forsythia (T.f.). The simultaneous quantitative determination of genome copies of P.g., T.d. and T.f. was demonstrated with a dynamic range from 0.2 to 1.5 x 10(5) unit/mu L. Results showed that this mPCR-qCE assay had run-to-run and day-to-day variations of 6.1-10.9% and 2.3-17.4%, respectively. As little as 4 pathogen cells would be sufficient to have a positive result. Finally, a clinical sample was determined and its P.g., T.d. and T.f. concentrations were 39720 unit/mu L, 5214 unit/mu L, and 1140 unit/mu L, respectively. This mPCR-qCE assay was proved to have comparable precision and sensitivity with quantitative PCR technology. Besides, it was more cost-effective, time saving and more capable of simultaneous detection. (C) 2017 Elsevier B.V. All rights reserved.