postheadericon N. meningitidis Serogroups

 Dia-MenSerogroup-020

Neisseria meningitidis (gram-negative bacteria) leads to meningitis, and is thus responsible for considerable mortality throughout the world. Most disease-causing Neisseira meningitidis strains belong to serogroups B and C in Europe, North & South America and Australia.

VALIDATION

Researched pathogens • Neisseria meningitidis (all strains): FAM dye, emission: 520 nm
• Serogroup A: FAM dye, emission: 520 nm
• Serogroup B: FAM dye, emission: 520 nm
• Serogroup C: FAM dye, emission: 520 nm
• Serogroup W135: FAM dye, emission: 520 nm
• Serogroup Y: FAM dye, emission: 520 nm
Target genes • CtrA (Neisseria meningitidis)
• sacB (Serogroup A)
• siaD (Serogroup B)
• siaD (Serogroup C)
• synG (Serogroup W135)
• synF (Serogroup Y)
Number of wells by test 6 (Neisseria meningitidis and serogroup A, B, C, W135 and Y)
Samples used • Human cerebrospinal fluid
• Whole blood
Validated extraction methods • Nuclisens easyMAG® System (Biomerieux)
• QIAamp® DNA Mini Kit (50) (Blood and Body Fluid Spin Protocol) (Qiagen)
• MagNa pure LC system (Roche) (Ongoing)
Real-time PCR systems • ABI (7000/7300/7500/7900)
• Roche LightCycler® (2.0/480)
• Biorad (iCycler/IQ5/CFX96/Dx)
• 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 nm
• DIA-UIC(DR)-050: orange dye, emission: 576 nm
• DIA-UIC(TR)-050: texas red dye, emission: 603 nm
• DIA-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 nm
• DIA-EIC/DNA(DR)-050: orange dye, emission: 576 nm
• DIA-EIC/DNA(TR)-050: texas red dye, emission: 603 nm
• DIA-EIC/DNA(Cy5)-050: Cy5 dye, emission: 662 nm