Saliva sampling methods. Cariogenic streptococci count using two different methods of saliva collection in children.

Abstract

The aim of this study is to compare the efficacy of two methods for collecting saliva samples from infants under 2 years of age for cariogenic streptococci (CS) count. Two collection methods were applied in 11 infants. In Method (A), saliva samples were collected by swabbing the inner cheek mucosa and floor of the mouth in figure of eight motions with a sterile cotton swab until it was soaked. In method (B), saliva samples were collected by aspiration of 1 ml of saliva with a sterile plastic syringe on the floor of the mouth, after stimulation with glove. The samples were cultured in modified Gold's broth (MSMG), and on trypticase, yeast extract, sucrose, cystine and bacitracin culture medium (TYSCB). In method (A), the swab with the sample was unloaded in situ on TYSCB and placed in PBS medium for transport. Then, 100 μl of the eluate was seeded in MSMG. In method (B) 100 μl were seeded in TYSCB and 100 μl in MSMG. Both culture media were incubatedundercapnophilicconditions for 48 hours at 37 °C. Colony forming units (CFU/ml) were counted by calibrated operators (kappa = 0.75). The presence of cariogenic streptococci (CS) (Streptococcus mutans-Streptococcus sobrinus) was determined by qPCR in the samples collected by both methods. The CFU/ml counts in MSMG differed significantly between methods (p = 0.021). In TYSCB, the recovery of CFU/ml was higher in method (A), without significant difference (p = 0.705). The molecular technique detected presence of CS, with no difference between collection methods. Collecting saliva samples by swabbing proved more effective in terms of recovery of microorganisms, and did not affect the detection of presence of CS by molecular techniques.