Human Infectiology MDx | Catalogue 2011 - Diagenode Diagnostics

HUMAN MOLECULAR DIAGNOSTICS CATALOGUE 2011RESPIRATORY INFECTIONSBordetella multiplexRef: Dia-Bor-050Whooping-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: yellow dye, emission: 549 nm• IS481 (Bordetella pertussis)• IS1001 (Bordetella parapertussis)Number of wells by test: 1 (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)• Amplicor respiratory specimen preparation kit (RSP prep) (Roche)Real-time PCR systems: • ABI (7000/7300/7500/7900)• Roche LightCycler ® (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:• 50 reactions (50 µl PCR, final volume)• 100 reactions (25 µl PCR, final volume)In combination with:• Universal inhibition control (DIA-UIC-050):DIA-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(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 nm26 | www.diagenodediagnostics.com