Bacterial Enteric Pathogens, Molecular Detection, PCR, Feces
Method Description Describes how the test is performed and provides a method-specific reference
This method employs a target-specific amplification and detection system using target specific primers in combination with fluorescent resonance energy transfer (FRET) hybridization probes designed for invA of Salmonella species, iphA found in both Shigella species and enteroinvasive Escherichia coli, cadF of Campylobacter jejuni and Campylobacter coli, stx1 and stx2 of Shiga toxin-producing Escherichia coli, and lysP of Yersinia species. The LightCycler instrument amplifies and monitors target nucleic acid sequences by fluorescence during PCR cycling. This is an automated PCR system that can rapidly detect amplified product development through stringent air-controlled temperature cycling and capillary cuvettes. The detection of amplified products is based on the FRET principle. For FRET product detection, a hybridization probe with a donor fluorophore, fluorescein, on the 3' end is excited by an external light source, which emits light that is absorbed by a second hybridization probed with an acceptor fluorophore, LC-Red 610, at the 5' end. The acceptor fluorophore then emits a light of a different wavelength that can be measured with a signal that is proportional to the amount of specific PCR product. For added specificity, probes are re-annealed to the amplified product and slowly disassociated by a graduated temperature increase. This process generates a melting curve and an associate melting temperature, specific to the assay target. The detection process is completed in less than 1 hour using a closed tube system.(Cunningham SA, Sloan LM, Nyre LM, et al: Three-hour molecular detection of Campylobacter, Salmonella, Yersinia, and Shigella species in feces with accuracy as high as that of culture. J Clin Microbiol 2010 Aug;48:2929-2933)
Supplemental Report Indicates whether the report includes an additional document with charts, images or other enriched information
Day(s) and Time(s) Test Performed Outlines the days and times the test is performed. This field reflects the day and time the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time required before the test is performed. Some tests are listed as continuously performed, which means assays are performed several times during the day.
Monday through Sunday; Once daily
Analytic Time Defines the amount of time it takes the laboratory to setup and perform the test. This is defined in number of days. The shortest interval of time expressed is "same day/1 day," which means the results may be available the same day that the sample is received in the testing laboratory. One day means results are available 1 day after the sample is received in the laboratory.
Maximum Laboratory Time Defines the maximum time from specimen receipt at Mayo Medical Laboratories until the release of the test result
Specimen Retention Time Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded
Performing Laboratory Location The location of the laboratory that performs the test