Bordetella pertussis and Bordetella parapertussis, Molecular Detection, PCR
Method Description Describes how the test is performed and provides a method-specific reference
The LightCycler instrument amplifies (multiplex) and monitors the development of target nucleic acid sequences by fluorescence after each cycle of PCR. This is an automated PCR system that can detect amplicon development through stringent air-controlled temperature cycling and cuvettes in approximately 60 minutes. The detection of amplified products is based on the fluorescence resonance energy transfer (FRET) principle. The assay uses the repetitive (50-100 copies) insertion sequence (IS481) found in Bordetella pertussis and the repetitive (35-50 copies) insertion sequence (IS1001) found in Bordetella parapertussis as targets. Detection and differentiation of Bordetella targets is performed through melting curve analysis. The probes were designed to obtain a 10 degrees C temperature shift between Bordetella pertussis and Bordetella parapertussis that is seen in the melting curve analysis. Analysis of the PCR amplification and probe melting curves is accomplished through the use of the LightCycler software. (Sloan LM, Hopkins MK, Mitchell PS, et al: Multiplex LightCycler PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussis in nasopharyngeal specimens. J Clin Microbiol 2002;40:96-100)
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 Friday; 10 a.m., Sunday; 10 a.m.
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