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. 2016 May 18;7:753. doi: 10.3389/fmicb.2016.00753

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Copyright © 2016 Wang, Wang, Zhang, Liu, Luo, Li, Cao, Liu, Xu and Ye.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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The sensitivity of multiplex ET-MCDA approach for simultaneously detecting two target pathogens in artificially contaminated blood samples. Two sets of ET-MCDA primers targeting invA and ipaH genes were simultaneously added to a reaction vessel. (A,B) were simultaneously yielded from Cy5 (labeling Sal-E-CP1 of invA) and Hex (labeling Shi-E-CP1 of ipaH) channels, respectively. Sensitivity of multiplex-ET-MCDA for simultaneously detecting Salmonella (A) and Shigella (B) in artificially contaminates blood samples was monitored by real-time format, and signals 1, 2, 3, 4, 5, 6, 7, and 8 represent Salmonella DNA levels of 39,000, 3900, 390, 39, 3.9, 0.39, and 0.039 CFU per reaction and negative control; Salmonella DNA levels for 34,000, 3400, 340, 34, 3.4, 0.34, and 0.034 CFU per reaction and negative control. The LoD of multiplex ET-MCDA method for Salmonella analysis in artificially contaminates blood samples was 3.9 CFU per reaction, and the LoD of multiplex ET-MCDA for Salmonella detection in artificially contaminates blood samples was 3.4 CFU per reaction.