Luminex is software

Blood samples were collected, incubated at room temperature, and centrifuged at 6,797 × g. Cytokines were quantified in sera by using a Luminex Screening assay (R&D Systems, Wiesbaden, Germany) and a Luminex 200 system with Luminex IS software (Luminex Corporation, MV’s-Hertogenbosch, Netherlands) according to the manufacturer’s instructions.

Blood samples were collected, incubated at room temperature and centrifuged at 1,000 × g. Splenic CD11c⁺ DCs were stimulated overnight with 100 ng/ml liposaccharide (Sigma-Aldrich, München, Germany). Cytokines were quantified in the supernatants from stimulated DCs or CD4⁺ T cells, and in sera by using a Luminex Screening assay (R&D Systems, Wiesbaden, Germany) and a Luminex 200 system with Luminex IS software (Luminex Corporation, MV’s-Hertogenbosch, Netherlands) according to the manufacturer’s instructions. The detection limits were 1.32 pg/ml for IL-10, 0.73 pg/ml for IFN-γ, 0.13 pg/ml for TNF-α, and 1.31 pg/ml for IL-12, respectively.

The extracted DNA was amplified by a nested PCR reaction using PGMY09/PGMY11 and GP5+/GP6+ primers as previously described [30 (link)].
The typing of 46 mucosal HPV types was carried out by a multiplex system based on the xMAP® technology as previously described by Zubach et al., [30 (link)]. Briefly, 46 fluorescence sortable microspheres (Luminex Corporation, Austin, TX) were coupled to the 46 specific probes for HPV types 6, 11, 13, 16, 18, 26, 30, 31, 32, 33, 34, 35, 39, 40, 42, 43, 44, 45, 51, 52, 53, 54, 56, 58, 59, 61, 62, 66, 67, 68, 69, 70, 71, 72, 73, 74, 81, 82, 83, 84, 85, 86, 87, 89, 90 and 91.. The double stranded second round PCR products, labelled with biotin, were made single-stranded by digestion with 2 μL of bacteriophage T7 gene6 exonuclease (New England BioLabs, Pickering, ON, Canada) that removed the non-labelled strand after 40 min incubation at room temperature. The single stranded HPV DNA was incubated for hybridization at 60 °C for 10 min and streptavidin-phycoerythrin (PE) (Invitrogen) in 1-tetramethyl ammonium chloride (TMAC) (Sigma), was added and incubated for 5 min at 60 °C. Genotype specific hybridization was detected on a Luminex Liquid Chip 200 flow cytometer (Qiagen) using the Luminex IS software (Luminex).