Specific primers for detection of Alicyclobacillus acidoterrestris by RT-PCR

The reverse transcription-polymerase chain reaction (RT-PCR), after a short enrichment culture, was used to detect Alicyclobacillus acidoterrestris in acidic beverages. Two specific primers were selected from the region of V2 and V4 on 16S rRNA gene. With this primer set, 294-bp fragments from ii. acidoterrestris could be amplified. The detection limit of the RT-PCR with the FHLP filters was about 10(-1) fg of pure total RNA per reaction. Juice samples inoculated with 10(4) cfu of A. acidoterrestris per mi were RT-PCR positive without enrichment. However, after 15 h of enrichment, the samples inoculated with 2-3 cfu ml(-1) were positive. This RT-PCR culture assay would enable rapid and specific detection of strains of A. acidoterrestris in acidic beverages.