O Serogroup-Specific Touchdown-Multiplex Polymerase Chain Reaction for Detection and Identification of Vibrio cholerae O1, O139, and Non-O1/Non-O139

Figure 3

Agarose gel electrophoresis analysis of the amplification efficiency of semiquantitative TMPCR. A reciprocal amplification algorithm of detectable levels of the O1 to O139 gDNAs (experiments (a) to (e)) was described in the text, while E. coli gDNA template serving as negative control (NC) was included. (a) Lanes 1 to 6, the 100 ng O1 gDNA versus a 10-fold serially diluted O139 gDNA, 1 pg to 100 ng, and vice versa (lanes 7 to 12). (b) Lanes 1 to 3, unequal amount ratios of the O1 : O139, 100 ng to 1 pg, 10 ng to 10 pg, and 1 ng to 100 pg, and vice versa (lanes 4 to 6). Lanes 7 to 12, the 100 pg O139 gDNA versus the O1 gDNA contents of varying 100 ng, 50 ng, 25 ng, 10 ng, and 1 ng to 100 pg. (c) Lanes 1 to 6, the equal amount ratios of 10-fold serially diluted O1 to O139 gDNAs, 100 ng to 1 pg. (d) Lanes 1 to 6, 10-fold serially diluted O1 gDNA contents, 100 ng to 1 pg. (e) Lanes 1 to 6, 10-fold serially diluted O139 gDNA contents, 100 ng to 1 pg. A 100 bp DNA ladder marker (lane M) was used in comparison of the amplicons with expected size (bp).

(a)

(b)

(c)

(d)

(e)