Multiplex qPCR for Meningococcal Genogroups

Two microliters of DNA extracted from culture-enriched samples were tested in 12.5 µl of reaction volume in qPCRs targeting genogroups A, B, C, W or Y^{24 (link)}. Primer and probe concentrations are listed in Table S1. These qPCRs were conducted on a LightCycler480, using SensiFast probe No-ROX mastermix (Bioline, London, United Kingdom) and with programme described in Table S2. Culture-enriched samples were regarded as positive for a genogroup when the C_T was lower than the cut-off value set for metA and ctrA. Control strains are listed in Table S3.

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Miellet W.R., Mariman R., Pluister G., de Jong L.J., Grift I., Wijkstra S., van Logchem E.M., van Veldhuizen J., Immink M.M., Wijmenga-Monsuur A.J., Rots N.Y., Sanders E.A., Bosch T, & Trzciński K. (2021). Detection of Neisseria meningitidis in saliva and oropharyngeal samples from college students. Scientific Reports, 11, 23138.

Publication 2021

LightCycler480 SensiFast probe No-ROX mastermix

Genogroup Primer Strains

Corresponding Organization :

Other organizations : National Institute for Public Health and the Environment, University of Applied Sciences Utrecht, Wilhelmina Children's Hospital, University Medical Center Utrecht

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Variable analysis

independent variables

Genogroups targeted (A, B, C, W or Y)

dependent variables

C_T (cycle threshold) values for the targeted genogroups

control variables

Reaction volume (12.5 µl)
DNA input (2 µl of extracted DNA from culture-enriched samples)
Primer and probe concentrations (listed in Table S1)
Thermal cycling program (described in Table S2)
Instrument (LightCycler480)
Mastermix (SensiFast probe No-ROX)

controls

Positive controls (control strains listed in Table S3)
Negative controls (not explicitly mentioned)

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