US 2014/0005061 A1 · COMPOSITIONS AND METHODS FOR DETECTION OF MULTIPLE MICROORGANISMS

Abstract

The present disclosure describes compositions, methods and kits for detection of one or multiple microorganism contaminants in samples. Some embodiments relate to detecting one or more microorganisms producing virulence factors such as shiga toxin stx1 and stx2 and eae. Some embodiments relate to detection of STEC microorganisms including an E. coli O26, an E. coli O45, an E. coli O103, an E. coli O111, an E. coli O121 or an E. coli O145. In some embodiments, compositions, methods and kits can detect and identify individual serotypes of shiga toxin producing microorganisms. Workflows for multiple microbe detection and identification are also described.

Claims21

Dependent claims

An isolated nucleic acid sequence comprising SEQ ID NO:1-SEQ ID NO:32 and SEQ ID NO:

39-SEQ ID NO:
203, fragments thereof, complements thereof, sequences comprising at least 90% nucleic acid sequence identity thereto and labeled derivatives thereof.

A method for detecting a shiga toxin producing E. Coli (STEC) microorganism in a sample comprising:

hybridizing at least a first pair of primers to a STEC target nucleic acid, a fragment thereof, a complements thereof, an allele thereof, or a variant thereof, to at least a first target polynucleotide sequence present in the sample;
amplifying at least the first target polynucleotide sequence or a fragment or a complement thereof to obtain at least one amplified target polynucleotide sequence;
detecting the at least one amplified target polynucleotide sequence;
and
the method optionally further comprising using a probe to detect the at least one amplified target polynucleotide sequence,
wherein detection of the at least one amplified target polynucleotide sequence is indicative of the presence of an STEC microorganism in the sample, wherein the STEC microorganisms is an E. coli O157:H7, an E. coli O26, an E. coli O45, an E. coli O103, an E. coli O111, an E. coli O121 or an E. coli O145.

3 more elements

A method for detection of one or more microorganisms expressing at least one virulence factor comprising:

1) detecting the presence of one or more virulence factors comprising:
a) contacting a sample suspected of containing one or more microorganisms expressing at least one virulence factors with one or more sets of primers specific to the one or more virulence factors;
b) amplifying at least one target nucleic acid sequence encoding at least one of the virulence factors to obtain one or more amplified virulence factor specific nucleic acids;
c) detecting the one or more amplified virulence factor specific nucleic acids or fragments or complements thereof;
and
d) optionally identifying the amplified virulence factor specific nucleic acids,
wherein detecting the one or more amplified virulence factor specific nucleic acids or fragments or complements thereof is indicative of the presence of a microorganism in the sample expressing the at least one virulence factor;
and
2) determining strain of the one or more microorganism expressing the at least one virulence factor comprising:
a) contacting the sample with at least one set of primers specific to a strain of the one or more microorganism;
b) amplifying at least one strain specific target nucleic acid sequence encoding for nucleic acid targets specific to the at least one strain of the microorganism to obtain one or more amplified strain specific nucleic acids;
c) detecting the one or more amplified strain specific nucleic acid or fragments or complements thereof;
and
d) optionally identifying the amplified strain specific nucleic acids.

11 more elements

Dependencies are inferred from the claim text.

Compositions and methods for detection of multiple microorganisms

Life of this patent

Compositions and methods for detection of multiple microorganisms

Life of this patent

Abstract

Claims21