PMID- 22916884
OWN - NLM
STAT- MEDLINE
DCOM- 20130201
LR  - 20221207
IS  - 1525-3198 (Electronic)
IS  - 0022-0302 (Linking)
VI  - 95
IP  - 9
DP  - 2012 Sep
TI  - Strain-level characterization of nonstarter lactic acid bacteria in Norvegia 
      cheese by high-resolution melt analysis.
PG  - 4804-4812
LID - S0022-0302(12)00494-8 [pii]
LID - 10.3168/jds.2012-5386 [doi]
AB  - The nonstarter lactic acid bacteria (NSLAB) constitute an important microbial 
      group found during cheese ripening and they are thought to be fundamental to the 
      quality of cheese. Rapid and accurate diagnostic tests for NSLAB are important 
      for cheese quality control and in understanding the cheese ripening process. 
      Here, we present a novel rapid approach for strain-level characterization through 
      combined 16S rRNA gene and repetitive sequence-based high-resolution melt 
      analysis (HRM). The approach was demonstrated through the characterization of 94 
      isolates from Norvegia, a Gouda-type cheese. The HRM profiles of the V1 and V3 
      variable regions of the 16S rRNA gene of the isolates were compared with the HRM 
      profiles of 13 reference strains. The HRM profile comparison of the V1 and V3 
      regions of the 16S rRNA gene allowed discrimination of isolates and reference 
      strains. Among the cheese isolates, Lactobacillus casei/paracasei (62 isolates) 
      and Lactobacillus plantarum/Lactobacillus pentosus (27 isolates) were the 
      dominant species, whereas Lactobacillus curvatus/Lactobacillus sakei were found 
      occasionally (5 isolates). The HRM profiling of repetitive sequence-based PCR 
      using the (GTG)(5) primer was developed for strain-level characterization. The 
      clustering analysis of the HRM profiles showed high discriminatory power, similar 
      to that of cluster analysis based on the gel method. In conclusion, the HRM 
      approach in this study may be applied as a fast, accurate, and reproducible 
      method for characterization of the NSLAB microflora in cheese and may be 
      applicable to other microbial environments following selective plate culturing.
CI  - Copyright (c) 2012 American Dairy Science Association. Published by Elsevier Inc. 
      All rights reserved.
FAU - Porcellato, D
AU  - Porcellato D
AD  - Department of Chemistry, Biotechnology and Food Science, Norwegian University of 
      Life Science, PO Box 5003, N-1432 Aas, Norway. Electronic address: 
      davide.porcellato@umb.no.
FAU - Ostlie, H M
AU  - Ostlie HM
AD  - Department of Chemistry, Biotechnology and Food Science, Norwegian University of 
      Life Science, PO Box 5003, N-1432 Aas, Norway.
FAU - Liland, K H
AU  - Liland KH
AD  - Department of Chemistry, Biotechnology and Food Science, Norwegian University of 
      Life Science, PO Box 5003, N-1432 Aas, Norway.
FAU - Rudi, K
AU  - Rudi K
AD  - Department of Chemistry, Biotechnology and Food Science, Norwegian University of 
      Life Science, PO Box 5003, N-1432 Aas, Norway.
FAU - Isaksson, T
AU  - Isaksson T
AD  - Department of Chemistry, Biotechnology and Food Science, Norwegian University of 
      Life Science, PO Box 5003, N-1432 Aas, Norway.
FAU - Skeie, S B
AU  - Skeie SB
AD  - Department of Chemistry, Biotechnology and Food Science, Norwegian University of 
      Life Science, PO Box 5003, N-1432 Aas, Norway.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Dairy Sci
JT  - Journal of dairy science
JID - 2985126R
RN  - 0 (RNA, Ribosomal, 16S)
SB  - IM
MH  - Cheese/*microbiology
MH  - DNA Fingerprinting
MH  - Food Handling
MH  - Lactobacillus/genetics/*metabolism
MH  - Lacticaseibacillus casei/genetics/metabolism
MH  - Lactobacillus plantarum/genetics/metabolism
MH  - Nucleic Acid Denaturation
MH  - Phylogeny
MH  - Polymerase Chain Reaction
MH  - RNA, Ribosomal, 16S/genetics
MH  - Sequence Analysis, RNA
EDAT- 2012/08/25 06:00
MHDA- 2013/02/05 06:00
CRDT- 2012/08/25 06:00
PHST- 2012/01/26 00:00 [received]
PHST- 2012/05/10 00:00 [accepted]
PHST- 2012/08/25 06:00 [entrez]
PHST- 2012/08/25 06:00 [pubmed]
PHST- 2013/02/05 06:00 [medline]
AID - S0022-0302(12)00494-8 [pii]
AID - 10.3168/jds.2012-5386 [doi]
PST - ppublish
SO  - J Dairy Sci. 2012 Sep;95(9):4804-4812. doi: 10.3168/jds.2012-5386.