Universidad de Buenos Aires, Facultad de Odontología, Cátedra de Odontología Preventiva y Comunitaria, Buenos Aires, Argentina.
Universidad de Buenos Aires, Facultad de Odontología, Cátedra de Microbiología y Parasitología, Laboratorio de Diagnóstico Microbiológico, Buenos Aires, Argentina.
Acta Odontol Latinoam. 2022 Apr 30;35(1):51-57. doi: 10.54589/aol.35/1/51.
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.
本研究旨在比较两种收集 2 岁以下婴儿唾液样本用于检测致龋链球菌(CS)的方法的效果。在 11 名婴儿中应用了两种收集方法。在方法(A)中,用无菌棉拭子以 8 字形的方式擦拭内颊粘膜和口腔底部,直到拭子被浸湿,从而收集唾液样本。在方法(B)中,通过刺激手套后用无菌塑料注射器从口腔底部抽吸 1ml 唾液,收集唾液样本。将样品接种于改良 Gold's 肉汤(MSMG)和胰蛋白酶、酵母提取物、蔗糖、半胱氨酸和杆菌肽培养(TYSCB)培养基上。在方法(A)中,将带有样本的拭子就地卸载到 TYSCB 上,并置于 PBS 培养基中进行运输。然后,将 100 μl 洗脱液接种于 MSMG 中。在方法(B)中,将 100 μl 接种于 TYSCB 中,100 μl 接种于 MSMG 中。两种培养基均在 37°C 下的二氧化碳环境中孵育 48 小时。由经过校准的操作人员(kappa = 0.75)计数形成菌落的单位(CFU/ml)。通过两种方法收集的样本均通过 qPCR 确定致龋链球菌(CS)(变形链球菌-远缘链球菌)的存在。MSMG 中的 CFU/ml 计数在两种方法之间差异有统计学意义(p = 0.021)。在 TYSCB 中,方法(A)中的 CFU/ml 回收率更高,但差异无统计学意义(p = 0.705)。分子技术检测到 CS 的存在,两种收集方法之间无差异。拭子法收集唾液样本在微生物回收方面更有效,且不影响分子技术检测 CS 的存在。