Juice analysis with real-time PCR

GEN-IAL developed a multiplex real-time PCR method to detect and differentiate Alicyclobacillus species in fruit juices and concentrates.

Alicyclobacillus is a gram-positive, strictly aerobic, acidophilic and thermophilic spoilage bacteria, especially important for the fruit juice industry, where it has been shown to produce high amounts of guaiacol imparting off-flavors to fruit juices.

Alicyclobacillus is of special interest to the fruit juice canning industry because common pasteurization techniques do not deactivate the spores. Therefore, early screening of Alicyclobacillus is very important for the industry since contamination can lead to considerable loss of quality and subsequently to economic losses for the producers. Alicyclobacillus acidoterrestris is considered the most important spoilage species within the Alicyclobacillus genera, but other e.g. Alicyclobacillus acidocaldarius have also been isolated from spoiled products.

General culture methods take 10-12 days to detect the presence of Alicyclobacillus species in samples, further 24h to confirm guaiacol production and identification of isolates require even longer. To overcome this difficulty, GEN-IAL developed a multiplex real-time PCR method to detect and differentiate Alicyclobacillus species in fruit juices and concentrates much earlier:

First-Alicyclobacillus multiplex PCR kit  (Art. No. TPABM 0050)

1. one specific system for the detection of Alicyclobacillus acidoterrestris (FAM/DQ).
2. one specific system for the detection of Alicyclobacillus acidocaldarius (Cy5/DQ).
3. one specific system for the detection of the genera Alicyclobacillus spp. (HEX/DQ).

To avoid false negative PCR-results due to inhibitory components in the samples, an internal control (ROX/DQ) is amplified together in the same reaction vessel with the Alicyclobacillus specific sequence.