Development of a DNA Array for the Simultaneous Detection and Identification of Sugar Thick Juice Bacterial Contaminants

Abstract

Despite the use of generally accepted good storage practices, sugar thick juice degradation caused by microbiological contamination occasionally occurs, causing considerable financial loss. In this study, a DNA array was developed for simultaneous detection and identification of the most prominent microflora present during thick juice storage, which may cause degradation of the thick juice. Specific oligonucleotides were developed for several bacterial taxa, including the genera Bacillus, Kocuria, Staphylococcus and Tetragenococcus and the species Aerococcus viridans, Leuconostoc mesenteroides and Tetragenococcus halophilus. The DNA array was validated using both pure cultures and industrial samples. In addition, comparisons were made between the developed array, PCR assays specifically targeting the thick juice contaminants and classical microbial platings. The array was found to be reliable and sensitive enough to detect and identify the target bacteria. In addition, the array was used to monitor the target microbial populations in thick juice during long-term storage and degradation. Results are discussed in relation to DNA stability in thick juice.