Application Guide - QIAxcel Advanced

QIAxcel Advanced Application Guide 10/2016 111 4 kV for 10 s, sample injection at 2 kV for 10 s and separation at 3.5 kV for 700 seconds. The QX Alignment Marker 15 bp/3 kb was run simultaneously with the samples, and the sizes were estimated by comparison with a control sample containing all eleven possible amplicons (Table 1). Following analysis with the QIAxcel ScreenGel® software, the operating and analysis software for the QIAxcel Advanced system, the results are displayed along with comprehensive reports including gel images, electropherograms and the result table. It is possible to customize the report according to different laboratory requirements. Results and discussion The analysis showed that the samples tested were positive for various combinations of O groups and virulence genes, as shown in Table 4 and Figures 1 and 2. Lane O-type eaeA stx1 stx2 ehxA 1 O26 – + – + 2 O26 – + + + 3 O45 – + – + 4 O103 – + – + 5 O103 – + + + 6 O111 + + + + 7 O111 + + – + 8 O121 – – + + 9 O145 + + – + 10 O145 + + + + 11 O157 + + + + Table 4. Result table showing the presence (+) or absence (–) of four virulence genes in seven E. coli serogroups. Figure 1. QIAxcel Advanced analysis of 11 isolates tested with 11-gene multiplex PCR. The PCR included the major E. coli virulence genes stx1, stx2, eae and ehxA, along with E. coli O-antigens from O26, O45, O103, O111, O121, O145 and O157 serogroups. Lane A12 contains a DNA ladder, which includes all 11 possible amplicons sizes and serves as a size marker. Amplicon sizes were verified by sequencing. O26 O26 O103 O45 O111 O111 O121 O145 O145 O157 O103 M 15 – 168 – 230 – 296 – 375 – 417 – 477 – 523 – 567 – 655 – 740 – 890 – 3000 – – ehxA – O111 – O157 – eae – O26 – stx2 – O145 – O121 – stx1 – O103 – O45 bp

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