PCR and DGGE Detection Limits for Wine Spoilage Microbes

  • L. Bester Department of Food Science, Stellenbosch University, Private Bag X1, Matieland 7602, South Africa
  • M. Cameron Department of Food Science, Stellenbosch University, Private Bag X1, Matieland 7602, South Africa
  • M. du Toit Institute for Wine Biotechnology, Department of Viticulture and Oenology, Stellenbosch University, Private Bag X1, Matieland 7602, South Africa
  • R.C. Witthuhn Department of Food Science, Stellenbosch University, Private Bag X1, Matieland 7602, South Africa
DOI: https://doi.org/10.21548/31-1-1396

Abstract

In this study the culture-independent technique, polymerase chain reaction (PCR)-denaturing gradient gel
electrophoresis (DGGE), was investigated for the early detection and identification of possible spoilage microbes
in wine. PCR and DGGE conditions were successfully optimised with the universal primers HDA1GC and HDA2,
the bacteria-specific primers WBAC1GC-WBAC2, and the yeast-specific primers NL1GC and LS2. PCR and DGGE
detection limits were determined for Lactobacillus plantarum, Pediococcus pentosaceus, Acetobacter pasteurianus
and Brettanomyces bruxellensis when inoculated into sterile saline solution (SSS) and white wine at 106 cfu/mL
respectively. PCR detection limits were more sensitive (101 to 102 cfu/mL) than DGGE detection limits (101 to 104 cfu/
mL), with the exception of B. bruxellensis, which had higher PCR and DGGE detection limits than the other reference
microbes. PCR-DGGE analysis was also used successfully to detect and identify Lb. plantarum, A. pasteurianus and
B. bruxellensis at a concentration of 108 cfu/mL as part of mixed populations in SSS and white wine. PCR detection
limits of 101 cfu/mL were determined for all three reference microbes in mixed populations. The DGGE detection
limits were higher for mixed populations when compared to single strains.

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Published
2016-12-09
Issue
Vol. 31 No. 1 (2010)
Section
Articles

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