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. 2020 Jun 17;11:1107. doi: 10.3389/fmicb.2020.01107

Figure 1.

Figure 1

Schematic overview of the culture-independent method for quantification of viable Campylobacter in raw milk. The method comprises the following steps. Raw milk is diluted 10-fold in precooled Brucella broth to enable quantitative bacterial centrifugation. After careful withdrawal of non-pelleted raw milk residues, the pellet is resuspended in 1 ml PBS for viable/dead differentiation according to Krüger et al. (2014). Subsequently, the DNA intercalating agent is applied to block DNA of dead bacteria from PCR amplification. After DNA extraction qPCR is performed targeting the 16S rRNA sequence in intact and putatively infectious units of thermophilic Campylobacter according to Pacholewicz et al. (2019).