Sensitive and rapid detection of Alicyclobacillus acidoterrestris using loop-mediated isothermal amplification

BACKGROUND: A loop-mediated isothermal amplification (LAMP) assay was developed for the rapid detection (within 2 h) of Alicyclobacillus acidoterrestris. The assay detected the species-specific DNA sequence of the 16S-23S rRNA internal transcribed spacer. RESULTS: The eight strains of A. acidoterrestris were successfully amplified, but six strains of other bacillus Acidocaldarius and 13 bacterial species other than bacillus Acidocaldarius were not. The sensitivity of the LAMP assay was at 4.50 x 10(-2) cfu per tube. This sensitivity is greater than that obtained by polymerase chain reaction (PCR) assay. The LAMP assay was examined further for its ability to detect A. acidoterrestris in juice samples. The results were compared with those of conventional PCR detection. CONCLUSION: Results indicate that the proposed LAMP assay is a rapid, specific and sensitive method for detecting A. acidoterrestris. As the amplification has been conducted under isothermal conditions, only a water bath or heating block is needed to maintain the required temperature. Thus, the method can be generalised and popularised easily in the future. (C) 2011 Society of Chemical Industry