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A real‐time polymerase chain reaction‐based method for rapid and specific detection of spoilage Alicyclobacillus spp. in apple juice
Author(s) -
Luo H.,
Yousef A.E.,
Wang H.H.
Publication year - 2004
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2004.01596.x
Subject(s) - food spoilage , library science , citation , pet food , food science , computer science , chemistry , biology , bacteria , genetics
Aims: To develop a real‐time PCR‐based rapid detection method for spoilage Alicyclobacillus spp. in juice products. Methods and Results: The squalene‐hopene cyclase‐encoding gene was targeted for primer‐and‐probe development. Gene fragments from representative strains were cloned, and PCR primers and probe were designed by DNA sequence comparison. Selected bacteria were examined for cross‐reactivity by the new method. Cells were serially diluted in apple juice and saline, and examined by the new method to establish detection sensitivity. Using the newly developed Taqman ® real‐time PCR‐based method, strains of Alicyclobacillus acidocaldarius and A. acidoterrestris were detected without cross reactivity with other common food‐borne micro‐organisms. Detection of <10 cells per PCR reaction from juice samples was accomplished within 3–5 h. Conclusion: This is the first reported real‐time PCR‐based detection method for Alicyclobacillus spp. and its application in juice products is demonstrated. Significance and Impact of the Study: As a favourable alternative for the laborious and time‐consuming culture‐ or biochemical characterization‐based techniques, the system has great potential for industrial applications from raw material screening to final product quality control.