FTD Respiratory pathogens 21 plus - Mikrogen

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18. Validation 18.1 Specificity FTD Respiratory pathogens 21 plus version 5/ March 2012 english FTD Respiratory pathogens 21 plus The BLAST (www.ncbi.nlm.nih.gov/blast) search shows that the selected primers and probes only detect the correspondent respiratory viruses and bacteria. The primer and probes of different viruses and bacteria are not cross-reacting. The targeted sequences for each pathogen are listed in the table below (table 7). Table 7: Target sequences of each pathogen Pathogen Target FluA Matrix gene (pos1) H1N1 segment 6 of NA FluB segment 8 NS1/NEP Rhino 5`untranslated Region (UTR) Cor43 nucleocapsid protein (N) gene Cor63 nucleocapsid protein (N) gene Cor229 nucleocapsid protein (N) gene HKU nucleocapsid protein (N) gene Para2 hemagglutinin-neuraminidase (HN) mRNA Para3 hemagglutinin-neuraminidase (HN) mRNA Para4 fusion protein gene Para1 hemagglutinin-neuraminidase (HN) mRNA HMPV A fusion glycoprotein (F) gene HMPV B fusion glycoprotein (F) gene Mpneu adhesin P1 HboV NP1 gene EV parts of domain IV and V PV 5`untranslated Region (UTR) RSVA nucleocapsid protein gene RSVB nucleoprotein mRNA AV hexon gene Saur sensor histidin kinase vick gene HIB BexA gene Spneu LytA gene Cpneu RNA polymerase beta chain gene - 26 -
There are only two exceptional cases. FTD Respiratory pathogens 21 plus version 5/ March 2012 english FTD Respiratory pathogens 21 plus The primers and probes for the detection of Rhino also detect EV, but not the other way around. If you get a positive signal for Rhino and EV the patient is EV positive. If you just detect Rhino the patient is Rhino positive. The primers and probes for the detection of HIB give also a positive signal for Haemophilus parahaemolyticus which is associated with paryngitis. The specificity was validated with different respiratory negative samples. These did not generate any positive signal with the primers and probes which are included in the FTD Resp 21 plus. For the specificity of the FTD Resp 21 plus assay we analyzed a range of positive material containing bacterial and viral pathogens. No other bacteria or viruses got detected by the six PPmixes only the mentioned exceptional cases. 18.2 Sensitivity The analytical detection limit was tested by using a tenfold dilution series of plasmid DNA of known concentration (1E+09-1E+00 copies/ml) and the FTD Resp 21 plus (table 8). In addition a 10 time measuring surrounding the plasmid detection limit was performed (table 9). Furthermore the detection limit was determined by using diluted positive patient material calculated with plasmid DNA of known concentration (1E+09-1E+06 copies/ml) and the FTD Resp 21 plus (table 8). - 27 -
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Page 3 and 4: 1. Identification of the manufactur
Page 7 and 8: 5. Contents FTD Respiratory pathoge
Page 9 and 10: 8. Principle of the method FTD Resp
Page 11 and 12: 11. Procedure FTD Respiratory patho
Page 15 and 16: Preparation of a 96 well plate for
Page 17 and 18: 12. Programming of the thermo cycle
Page 25: 17. Troubleshooting FTD Respiratory
Page 31 and 32: Pathogen FluA FTD resp+ H1N1 FTD Fl

FTD Respiratory pathogens 21 plus - Mikrogen

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