Elisa assay kit

Two cohorts of participants (n=169) were included in this study. For the first cohort, stimulated saliva samples were collected from 120 patients diagnosed with CP (n=63) and healthy controls (n=57). The mean age ± SD and (range) for the patients with periodontitis was 58.8 ± 12.7 (22–86), and for the healthy control group 42.3 ± 13.5 (24–77) years. The ratio of males/females were 31/32 in the periodontitis group and 22/35 in the healthy control group. The clinical diagnosis of CP was based on tooth sites with probing pocket depth ≥ 6 mm, bleeding on probing >30%, clinical attachment level ≥ 5 mm and radiographic evidence of bone loss. For the second cohort (n=49), stimulated saliva samples were collected from healthy controls and patients with different severity of periodontitis, based on radiographic evidence of marginal bone loss and sites bleeding on probing with pocket depth > 4mm, and healthy control group (n=10). For this cohort, the mean age ± SD (range) for patients with periodontitis was 69.5 ± 8.0 (51–83) and for the healthy group 64.4 ± 15.5 (31–81) years. The participants included in the control groups showed no marginal bone loss, probing depth ≤ 3.0 mm and bleeding on probing < 10%.
Briefly, all participants fasted 2 h prior to the clinical examination. Masticatory stimulation was induced by chewing paraffin tablets for 2 minutes before saliva was collected in pre-labelled 50 ml sterile falcon tubes. After collection, the saliva samples were immediately frozen at −20°C until further processing as previously described (26 (link)). The samples where then thawed and centrifuged at 500 x g for 10 min at 5°C and the supernatants were stored at −80°C until analysis.
Saliva samples were analyzed using the Human PLEK (pleckstrin) ELISA assay kit with a range of 0.156-10 ng/ml and sensitivity of 0.094 ng/ml (catalog # EH11196, Wuhan Fine Biotech Co, China). The ELISA assay was executed following the manufacturer’s protocol.