Simultaneous quantification of pathogenic Campylobacter and Salmonella in chicken rinse fluid by a flotation and real-time multiplex PCR procedure

Quantifying viable Vibrio parahaemolyticus and Listeria monocytogenes simultaneously in raw shrimp

“…Although the TaqMan primers and probes of this method have been used in previous real-time PCR research (Nordstrom et al 2007;Omiccioli et al 2009;Zhang et al 2015), in this study, the high specificity was reconfirmed on tests using 96 strains (Table 1) due to the addition of PMA sample treatment step and the adjustment of the PCR system, which might influence the specificity of reaction (Wolffs et al 2007;Zhang et al 2015). The amplification efficiencies of the new PCR assay were tested by constructing standard curves on artificially contaminated raw shrimp model without any sterilization pretreatments.…”

“…Firstly, the time for development of models was greatly shortened by this new assay, while it took at least 3-5 days by traditional plate-counting method (Ye et al 2013;Wang et al 2014;Liao et al 2015). Secondly, the high sensitive and specified TaqMan probe made this new assay have ability to precisely describe the behavior of bacteria and be free from interference of raw food matrixes and complex natural microbiota (Wolffs et al 2007;Ma et al 2014;Zhang et al 2015). Thirdly, since adding the PMA treatment, this assay solved the problem that the common molecular method is not suited to describe the inactivation behavior of bacteria .…”

Enumeration of Salmonella Bacteria in Food and Feed Samples by Real-Time PCR for Quantitative Microbial Risk Assessment

“…Two-step extraction strategies separate the target cells from a food matrix prior to isolation of DNA from the capture matrix. Two-step cell extraction can be done using flotation (34,37), paramagnetic beads (28,32), or filtration (38).…”

“…This method was used to quantify Yersinia enterocolitica in pork (34) and Campylobacter in a chicken rinse (35), as well as to separate living cells from DNA of dead cells (34). Recently, a quantitative multiplex real-time PCR for Campylobacter and Salmonella based on a rather lengthy flotation procedure was described (37). However, there are problems with variation and lack of robustness when this sample preparation method is used, especially when a large number of samples must be processed simultaneously.…”

Qualitative and quantitative methodologies for determination of airborne microorganisms at concentrated animal-feeding operations

“…in soil (Ibekwe and Grieve 2003;Ibekwe et al 2004), feces (Bono et al 2004), wash wastewater (Ibekwe et al 2002;Malorny et al 2007;Wolffs et al 2007), and food (Fortin et al 2001;Heller et al 2003;Ellingson et al 2004). While the detection limit for these pathogens is generally [10 2 c.f.u.…”