A single enzyme PCR-RFLP assay targeting V1-V3 region of 16S rRNA gene for direct identification of Alicyclobacillus acidoterrestris from other Alicyclobacillus species.
Alicyclobacillus acidoterrestris is considered to be one of the most important target microorganisms in quality control of heat-processed acidic foods. This species can cause spoilage by forming spores with very high heat resistance; however, no gas production or visible changes occur in the contaminated product during storage. Thus, the differentiation of A. acidoterrestris from other Alicyclobacillus species is of great importance. Τhe present study aims to find a rapid method for the identification of A. acidoterrestris. To achieve this, 78 Alicyclobacillus isolates were subjected to PCR-RFLP (restriction fragment length polymorphism). Specifically, PCR products of amplified V1-V3 region of the 16S rRNA gene were digested with restriction endonuclease HhaI. According to the obtained results, all A. acidoterrestris isolates showed similar restriction fragments of the 16S rRNA gene and different from A. acidocaldarius and A. hesperidum. In conclusion, a single enzyme PCR-RFLP assay was developed and showed rapid, inexpensive and direct identification of Alicyclobacillus isolates. The application of this method will be useful to identify this contaminant in fruit juices.
['Sourri|Patra|P|', 'Doulgeraki|Agapi I|AI|http://orcid.org/0000-0001-9125-8856', 'Tassou|Chrysoula C|CC|', 'Nychas|George-John E|GE|']
D056450:Alicyclobacillus D004269:DNA, Bacterial D005516:Food Microbiology D000067030:Fruit and Vegetable Juices D005838:Genotype D006358:Hot Temperature D006801:Humans D010802:Phylogeny D016133:Polymerase Chain Reaction D012150:Polymorphism, Restriction Fragment Length D012336:RNA, Ribosomal, 16S D013045:Species Specificity
Alicyclobacillus A. acidoterrestris New assay RFLP Species differentiation