Rapid salmonella serotyping assay

Abstract

Processes for the serotype specific detection and identification of one or more Salmonella serotypes are provided. A family of specific primers and probes are provided that allow screening of biological or environmental samples for robust, rapid, and reproducible detection and identification of one or more Salmonella serotypes in the sample.

1 . A process of specifically identifying  Salmonella  ser.  Enteritidis  in a sample comprising:
 performing a first stage assay by adding to a sample a plurality of SE primer sets operable to produce amplification products of SE target sequences, said primer sets each comprising a forward primer that hybridizes to a region flanking a target sequence under conditions suitable for a polymerase chain reaction, and a reverse primer that hybridizes to a different region of the corresponding target sequence under conditions suitable for a polymerase chain reaction, wherein each target sequence is a portion of a  Salmonella  gene;   detecting the presence of or absence of each of said amplification products by hybridization of a probe specific to said detected amplification product.   
     
     
         2 . The process of  claim 1  wherein said probe is comprises the sequence of at least one of SEQ ID NOs: 117, 118, 119, 120, 121, 122, or 123. 
     
     
         3 . The process of  claim 1  wherein at least one of said forward primers comprises the sequence of any of SEQ ID NOs: 107, 109, 111, 113, or 115. 
     
     
         4 . The process of  claim 1  wherein at least one of said reverse primers has the sequence of at least one of SEQ ID NOs: 108, 110, 112, 114, or 116. 
     
     
         5 . A process of serotyping  Salmonella  in a sample comprising:
 adding to a sample one or more O-grp-2 primer sets operable to produce an O-grp-2 amplification product of O-grp-2 target sequence;   said O-grp-2 target sequence comprising one or more O-grp-2 target sequences selected from the group consisting of O:6,14; O:11; O:16; O:35; O:50; O:17, O18; O21, O28; O30, O38; O40, O44; O47, O48; and O61;   each of said primer sets comprising a forward primer that hybridizes to a region flanking a respective target sequence under conditions suitable for a polymerase chain reaction, and a reverse primer that hybridizes to a different region of the target sequence under conditions suitable for a polymerase chain reaction; and   detecting the presence or absence of each of said amplification products by hybridization to a labeled probe specific to said O-grp-2 amplification product a labeled probe specific to said O-grp-2 amplification product.   
     
     
         6 . The process of  claim 5  wherein at least one of said forward primers comprises the nucleotide sequence of any one or more of SEQ ID NOs: 92, 93, 94, 95, or 96. 
     
     
         7 . The process of  claim 5  wherein at least one of said reverse primers comprises the nucleotide sequence of any one or more of SEQ ID NOs: 97, 98, 99, 100, or 101. 
     
     
         8 . The process of  claim 5  wherein at least one of said probes comprises the nucleotide sequence of any one or more of SEQ ID NOs: 102, 103, 104, 105, or 106. 
     
     
         9 . The process of  claim 5  wherein said detecting diagnoses  Salmonella  infection in a subject.