[Study on application of rep-PCR fingerprint in rapid identification of beer-spoilager].
The application potential of rep-PCR in typing beer-spoilage isolates was studied. The effects of different factors, including DNA templates and primers, on the quality and reproducibility of fingerprints were investigated. The CTAB protocol was shown to be the feasible method for DNA extraction. Primers BOXA1R and (GTG)5 were used in rep-PCR, and the PCR products were sequenced to identify strains isolated from two breweries. Rep-PCR fingerprint profiles were obtained by using GelCompar II software. Cluster analysis showed that the isolates belonging to Lactobacillus brevis, L. buchneri, L. casei/paracasei, L. plantarum are divided into 2 or 3 subgroups. In addition, the two rep-PCR fingerprint profiles complemented with each other in typing these isolates. Combining the similarity coefficient cut-off (SCC) of species, 9 unknown isolates were identified rapidly by using both fingerprint databases. The results indicate that rep-PCR is a simple, reliable and promising method for rapid identification of beer-spoilager.
Zhu
Lin-Jiang
LJ
The Key Laboratory of Industrial Biotechnology, Ministry of Education, Wuxi 214036, China.
Zheng
Fei-Yun
FY
Zhao
Ya-Zhou
YZ
Xing
Xiang-Nan
XN
Li
Qi
Q
Gu
Guo-Xian
GX
D001515:Beer / Q000382:microbiology*; D016000:Cluster Analysis; D016172:DNA Fingerprinting / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D030541:Databases, Genetic; D007778:Lactobacillus / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D013997:Time Factors
D004269:DNA, Bacterial