Application Guide - QIAxcel Advanced

114 QIAxcel Advanced Application Guide 10/2016 Conclusions • The method presented here can identify the seven major E. coli serogroups and the four main virulence factors in one reaction. • Separating multiplex PCR products with the QIAxcel Advanced system allows detailed, reliable and rapid analyses of complex samples containing many amplicons within a wide size range. • Due to its fast turnaround time and automatic data interpretation, the QIAxcel Advanced system is highly suited for post-PCR separation, visualization and data presentation, especially for multiplex PCR runs in a high-throughput setting. Peak Calling Result Table E.coli 11-multiplex PCR Plate |R0|E0 Pos Sample Info ehxA wzxO111 rfbO157 eae wzxO26 stx2 wzxO145 wbqEO121 wbqFO121 stx1 wzxO103 wzxO45 A1 E.coli-A-01 169 n/a n/a n/a 419 n/a n/a n/a 667 n/a n/a A2 E.coli-A-02 168 n/a n/a n/a 418 479 n/a n/a 664 n/a n/a A3 E.coli-A-03 169 n/a n/a n/a n/a n/a n/a n/a 666 n/a 947 A4 E.coli-A-04 169 n/a n/a n/a n/a n/a n/a n/a 667 759 n/a A5 E.coli-A-05 169 n/a n/a n/a n/a 481 n/a n/a 666 757 n/a A6 E.coli-A-06 169 231 n/a 376 n/a 479 n/a n/a 664 n/a n/a A7 E.coli-A-07 169 231 n/a 376 n/a n/a n/a n/a 666 n/a n/a A8 E.coli-A-08 169 n/a n/a n/a n/a 479 n/a 595 n/a n/a n/a A9 E.coli-A-09 169 n/a n/a 380 n/a n/a 529 n/a 667 n/a n/a A10 E.coli-A-010 169 n/a n/a 380 n/a 482 529 n/a 667 n/a n/a A11 E.coli-A-011 168 n/a 297 378 n/a 480 n/a n/a 667 n/a n/a Table 6. Based on the data presented in Figure 1and the peak calling instructions, a peak calling result table was generated. The table summarizes the experiment by reporting for each sample presence or absence of amplification products and their size estimates. References 1. Rangel, J.M., Sparling, P.H., Crowe, C., Griffin, P.M., and Swerdlow, D.L. (2005) Epidemiology of Escherichia coli O157:H7 outbreaks, United States, 1982–2002. Emerg. Infect. Dis. 11, 603–609. 2. Hussein, H.S. and Bollinger, L.M. (2005) Prevalence of Shiga toxin-producing Escherichia coli in beef cattle. J. Food. Prot. 68, 2224–2241. 3. Ferens, W.A. and Hovde, C.J. (2011) Escherichia coli O157:H7: Animal reservoir and sources of human infection. Foodborne Pathog. Dis. 8, 465–487. 4. Bai, J., Shi, X., and Nagaraja, T.G. (2010) A multiplex procedure for the detection of six major virulence genes in Escherichia coli O157:H7. J. Microbiol. Meth. 82, 85–89. 5. Bai, J., Paddock, Z.D., Shi, X., Li, S., An, B., and Nagaraja, T.G. (2012) Applicability of a multiplex PCR to detect the seven major Shiga toxin-producing Escherichia coli based on genes that code for serogroup-specific O-antigens and major virulence factors in cattle feces. Foodborne Pathog. Dis. 9, 541–548. 6. Fratamico, P.M., Briggs, C.E., Needle, D., Chen, C.Y., and DebRoy, C. (2003) Sequence of the Escherichia coli O121 O-antigen gene cluster and detection of enterohemorrhagic E. coli O121 by PCR amplification of the wzx and wzy genes. J Clin. Microbiol. 7, 3379–3383. A1 E.coli-A-01169 n/a n/a n/a 419 n/a n/a n/a 667 n/a n/a A2 E.coli-A-02168 n/a n/a n/a 418479 n/a n/a 664 n/a n/a A3 E.coli-A-03169 n/a n/a n/a n/a n/a n/a n/a 666 n/a 947 A4 E.coli-A-04169 n/a n/a n/a n/a n/a n/a n/a 667759 n/a A5 E.coli-A-05169 n/a n/a n/a n/a 481 n/a n/a 666757 n/a A6 E.coli-A-06169231 n/a 376 n/a 479 n/a n/a 664 n/a n/a A7 E.coli-A-07169231 n/a 376 n/a n/a n/a n/a 666 n/a n/a A8 E.coli-A-08169 n/a n/a n/a n/a 479 n/a 595 n/a n/a n/a A9 E.coli-A-09169 n/a n/a 380 n/a n/a 529 n/a 667 n/a n/a A10 E.coli-A-010169 n/a n/a 380 n/a 482529 n/a 667 n/a n/a A11 E.coli-A-011168 n/a 297378 n/a 480 n/a n/a 667 n/a n/a

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