Boston Probes, Inc.

Applera Corporation

Life Technologies Corporation

Heather P O'Keefe

Jens J Hyldig-Nielsen

Henrik Stender

Susan Rigby

This invention is related to methods pertaining to the determination of bacteria and yeast wherein the methods comprise: a) filtering a sample containing bacteria and yeast through at least one filter; b) treating the filter or filters with a medium which facilitates the growth of the bacteria and yeast; c) growing the bacteria and yeast on the filter or filters to thereby produce colonies or microcolonies of the bacteria and yeast; d) fixing the colonies or microcolonies to the filter or filters; e) treating each filter under suitable hybridization conditions with one or more enzyme labeled PNA probes wherein, each PNA probe comprises a probing nucleobase sequence complementary to a target sequence in the nucleic acid of the bacteria and/or yeast; f) removing excess enzyme labeled PNA probe from each of the filters; and g) determining enzyme activity remaining on each filter to thereby identify or enumerate colonies or microcolonies on the filter or filters.

Methods for the analysis of microcolonies of bacteria and yeast

This invention is related to novel probes, probe sets, methods and kits pertaining to the detection, identification and/or quantitation of yeasts and particularly(a.k.a.) an organism that spoils wine. Preferred probes for the detection of one or more species of thegenus comprise a probing nucleobase sequence, at least a portion of which is selected from the group consisting of: AGC-GGG-TCT-ATT-AGA (Seq. ID No. 1); CCA-GGT-GAG-GGT-CGC (Seq. ID No. 2); CGG-TTG-CCC-GAT-TTC (Seq. ID No. 3); TCG-CCT-TCC-TCC-TCT (Seq. ID No. 4); CGG-TCT-CCA-GCG-ATT (Seq. ID No. 5); CAC-AAG-ATG-TCC-GCG (Seq. ID No. 6); GCG-GGC-ACT-AAT-TGA (Seq. ID No. 7); CAT-CCA-CGA-GGA-ACG (Seq. ID No. 8); GTG-TAA-ACC-AGG-TGC (Seq. ID No. 9); ATG-GCT-CCC-AGA-ACC (Seq. ID No. 10) and GAC-AGA-ATC-GAA-GGG (Seq. ID No. 11). The probes, probe sets, methods and kits of this invention are particularly well suited for use in the analysis of yeast in wine, beer and liquor and in the monitoring of contamination of in-process product as well as the equipment and facilities used to manufacture product in wineries and breweries.

Methods for the detection, identification, and/or enumeration of yeast, particularly in wine

This invention is directed to a rapid and simple method for determining organisms and/or cells in a sample. Generally, the method is directed to the use of molecular probes to selectively stain the organisms and/or cells for determination wherein growth medium, fixative reagents and/or excess molecular probes need not be separated before a determination is made.

Methods for determining organisms not requiring the separation of fixative or excess probe

This invention is related to novel PNA probes, probe sets, methods and kits pertaining to the universal detection of bacteria and/or eucarya. Preferred universal probes for the detection of bacteria comprise a probing nucleobase sequence selected from the group consisting of CTG-CCT-CCC-GTA-GGA; TAC-CAG-GGT-ATC-TAA-T; CAC-GAG-CTG-ACG-ACA and CCG-ACA-AGG-AAT-TTC. Preferred universal probes for the detection of eucarya comprise a probing nucleobase sequence selected from the group consisting of ACC-AGA-CTT-GCC-CTC-C; GGG-CAT-CAC-AGA-CCT-G; TAG-AAA-GGG-CAG-GGA and TAC-AAA-GGG-CAG-GGA. The PNA probes, probe sets, methods and kits of this invention are particularly well suited for use in multiplex PNA-FISH assays wherein the bacteria and/or eucarya in a sample can be individually detected, identified or quantitated. Using exemplary assays described herein, the total number of colony forming units (CFU) of bacteria and/or eucarya can be rapidly determined.

Growing community of inventors

Lexington, MA, United States of America

Heather P O'Keefe