MacKay B J, Goodman H, Cox D, Grossbard B L, Iacono V J, Pollock J J
J Clin Microbiol. 1984 Jun;19(6):844-8. doi: 10.1128/jcm.19.6.844-848.1984.
The specificity of lysozyme determinations in human parotid and submandibular-sublingual salivas of two subjects was assessed by comparison of lysozyme concentrations in native acidified salivas with purified enzyme obtained by immunoadsorbent fractionation of the salivas. Lysozyme concentrations were measured by the turbidimetric catalytic method and by a newly developed enzyme-linked immunosorbent assay (ELISA). The validity of the assays was established by comparing assay results with enzyme concentration values determined from optical density-extinction coefficient calculations of the purified lysozyme peak. Values for purified enzyme were found to be similar, irrespective of the assay used to determine lysozyme concentrations, and were in agreement with extinction coefficient calculations. Based on the ELISA technique, recoveries of lysozyme from both parotid and submandibular-sublingual salivas were greater than 75 and 90%, respectively. Similar recoveries were noted for parotid saliva when determinations were based on the turbidimetric assay. However, the ELISA and turbidimetric assays differed with respect to lysozyme levels in submandibular-sublingual saliva because of the apparent presence of an enhancement factor which gave rise to higher lysozyme values in the catalytic assay and therefore resulted in low recoveries of purified enzyme. This catalytic enhancement factor was present in the nonadsorbed fraction of both subjects, as higher lysozyme activities were noted when nonadsorbed fractions were added to affinity-purified lysozymes. Lysozyme levels were also determined in the parotid and submandibular-sublingual salivas of caries-resistant and -susceptible adults. In general, levels of lysozyme in parotid saliva were lower in comparison to submandibular -sublingual saliva; however, significant differences in enzyme concentration were not evident between the caries-resistant and caries-susceptible subjects.
通过比较天然酸化唾液中的溶菌酶浓度与通过唾液免疫吸附分级分离获得的纯化酶，评估了两名受试者腮腺唾液和颌下-舌下唾液中溶菌酶测定的特异性。溶菌酶浓度通过比浊催化法和新开发的酶联免疫吸附测定法（ELISA）进行测量。通过将测定结果与根据纯化溶菌酶峰的光密度-消光系数计算得出的酶浓度值进行比较，确定了测定方法的有效性。无论使用何种测定方法来确定溶菌酶浓度，纯化酶的值都相似，并且与消光系数计算结果一致。基于ELISA技术，腮腺唾液和颌下-舌下唾液中溶菌酶的回收率分别大于75%和90%。当基于比浊法进行测定时，腮腺唾液的回收率相似。然而，ELISA法和比浊法在颌下-舌下唾液中的溶菌酶水平方面存在差异，因为明显存在一种增强因子，该因子在催化测定中导致溶菌酶值更高，因此导致纯化酶的回收率较低。这种催化增强因子存在于两名受试者的未吸附部分中，因为当将未吸附部分添加到亲和纯化的溶菌酶中时，观察到更高的溶菌酶活性。还测定了龋齿易感和抗龋齿成年人的腮腺唾液和颌下-舌下唾液中的溶菌酶水平。一般来说，腮腺唾液中的溶菌酶水平低于颌下-舌下唾液；然而，龋齿易感和抗龋齿受试者之间的酶浓度没有明显差异。
