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Simultaneous quantitative detection of  Lactococcus  ( Lc .)  lactis  and  Leuconostoc  species bacteriophages (phages) has not been reported in dairies using undefined mixed-strain DL-starters, probably due to the lack of applicable methods. We optimized a high-throughput qPCR system that allows simultaneous quantitative detection of  Lc .  lactis  936 (now  SK1virus ), P335, c2 (now  C2virus ) and  Leuconostoc  phage groups. Component assays are designed to have high efficiencies and nearly the same dynamic detection ranges,  i . e ., from ~1.1 x 10 5  to ~1.1 x 10 1  phage genomes per reaction, which corresponds to ~9 x 10 7  to ~9 x 10 3  phage particles mL -1  without any additional up-concentrating steps. The amplification efficiencies of the corresponding assays were 100.1±2.6, 98.7±2.3, 101.0±2.3 and 96.2±6.2. The qPCR system was tested on samples obtained from a dairy plant that employed traditional mother-bulk-cheese vat system. High levels of 936 and P335 phages were detected in the mother culture and the bulk starter, but also in the whey samples. Low levels of phages were detected in the cheese milk samples.

A high-throughput qPCR system for simultaneous quantitative detection of dairy Lactococcus lactis and Leuconostoc bacteriophages