Human Infectiology MDx | Catalogue 2011 - Diagenode Diagnostics

BordetellaRef: Dia-Bor-020Whooping-cough is a highly contagious respiratory illness causedby the gram-negative bacterial pathogen Bordetella pertussis(Cherry, 2004). Among the nine species of Bordetella identifiedtoday, only three additional members (B.parapertussis, B.holmesii,B.bronchiseptica) have been associated with respiratory infectionsin humans and others mammals.Cherry, J. D., and U.Heininger. (2004). Pertussis and other Bordetella infections.In R.D.Feigin, J.D.Cherry, G.J.Demmler, and S.Kaplan (ed.), Textbook ofpediatric infectious diseases, 1588-1608.VALIDATIONResearched pathogens:Target genes:• Bordetella pertussis: FAM dye, emission: 520 nm• Bordetella parapertussis: FAM dye, emission: 520 nm• IS481 (Bordetella pertussis)• IS1001 (Bordetella parapertussis)Number of wells by test: 2 (Bordetella pertussis and Bordetella parapertussis)Samples used:• Bronchoalveolar lavage• Sputum• Nasopharyngeal swabs/washesValidated extraction methods: • Nuclisens easyMAG ® System (Biomerieux)• QIAamp ® DNA Mini Kit (50) (Blood and Body Fluid Spin Protocol) (Qiagen)• MagNa pure LC system (Roche)• QiaSymphony (Qiagen)Real-time PCR systems: • ABI (7000/7300/7500/7900)• Roche LightCycler ® (2.0/480)• Biorad (iCycler/IQ5/CFX96)• Qiagen Rotor-Gene (6000)• Stratagene (MX3000P/3005P)• Cepheid (SmartCycler II)PCR volumes: 50 µl, 25 µl, 20 µl (depending of the real-time PCR system used)PCR reactions/kit:• 20 reactions (50 µl PCR, final volume)• 40 reactions (25 µl PCR, final volume)• 50 reactions (20 µl PCR, final volume)In combination with:• Universal inhibition control (DIA-UIC-050):DIA-UIC(YD)-050: yellow dye, emission: 549 nmDIA-UIC(DR)-050: orange dye, emission: 576 nmDIA-UIC(TR)-050: texas red dye, emission: 603 nmDIA-UIC(Cy5)-050: Cy5 dye, emission: 662 nm• DNA extraction & PCR inhibition control (DIA-EIC/DNA)-050:DIA-EIC/DNA(YD)-050: yellow dye, emission: 549 nmDIA-EIC/DNA(DR)-050: orange dye, emission: 576 nmDIA-EIC/DNA(TR)-050: texas red dye, emission: 603 nmDIA-EIC/DNA(Cy5)-050: Cy5 dye, emission: 662 nmRESPIRATORY INFECTIONS25