Human Infectiology MDx | Catalogue 2011 - Diagenode Diagnostics

Mycoplasma pneumoniae &Chlamydophila pneumoniaeRef: Dia-MCpn-050Mycoplasma pneumoniae and Chlamydophila pneumoniae arethe organisms responsible for most of the cases of atypicalpneumonia in childrenAtypical pneumonia due to these bacteria usually causes mildforms of pneumonia and are characterized by a more protractedcourse of symptoms unlike other forms of pneumonia which canprogress more quickly with more severe early symptoms.Mycoplasma pneumoniae can give rise to mild upper respiratorytract infection, bronchitis, bronchiolitis and bronchopneumonia.The disease usually starts as influenza like syndrome withfever, illness, headache, scratchy sore throat and cough as thepredominant symptoms.Chlamydophila pneumoniae has been associated with pharyngitis,sinusitis and bronchitis.VALIDATIONResearched pathogens:Target genes:• Mycoplasma pneumoniae: FAM dye, emission: 520 nm• Chlamydophila pneumoniae: yellow dye, emission: 549 nm• P1 (Mycoplasma pneumoniae)• 16S (Chlamydophila pneumoniae)Number of wells by test: 1 (Mycoplasma pneumoniae / Chlamydophila pneumoniae)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)• MagNa pure LC system (Roche)• QiaSymphony (Qiagen)Real-time PCR systems: • ABI (7000/7300/7500/7900)• Roche LightCycler ® (480/2.0 on request)• Biorad (iCycler/IQ5/CFX96)• Qiagen Rotor-Gene (6000)• Stratagene (MX3000P/3005P)• Cepheid (SmartCycler II)PCR volumes: 50 µl, 25 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 nm35