Bozhko N V, Lebedeva S A, Bichul' O K, Ivanova V S, Lysova L K, Barabash G P, Nekliaev V N, trukhachev A L, Terent'ev A N
Klin Lab Diagn. 2005 Jun(6):45-9.
The experimentally obtained antigenic complex isolated by extraction in the gradient surfactants from live and acetone-dried bacteria of the capsule-free vaccine strain EV76 of a plague microbe that had lost its ability to synthesize the diagnostic species-specific capsular antigen F1 was investigated. The antigenic complex fraction V (FV) was obtained after the fifth stage of extraction at a concentration of 1.28% of surfactants and after additional purification. The thermostable FV was found to consist mainly of protein. The protein having a molecular mass of about 43 kD predominates in the fraction. The latter is nontoxic for albino mice and antigenic. It forms a precipitate with commercial antiplague serum antibodies. FV antigenic sensitization of tanned sheep red blood cells gave rise to a diagnostic agent that specifically reacted with an antiplague serum rather than with heterologous sera against enterobacteria. The sera immunized with FV specifically reacted in the JDJFR with all the strains of the pathogen of plague irrespective of the temperature of their cultivation, including "fraction-free", which did not interact with a diagnosticum on F1. The animal sera immunized with capsule-free plague microbial strain reacted only with a FV-erythrocytic diagnosticum and they did not interact with F1 antigen-sensitized red blood cells. The erythrocytic FV diagnosticum was tested in ABNR with 130 typical and atypical plague microbial strains and with 133 strains of heterologous bacteria of different species of the family Enterobacteriaceae. The FV diagnosticum identified all the variants of a plague microbe, while the F1 diagnosticum revealed only its capsular variants. Among the heterologous bacteria, some strains of the closely related pathogen of pseudotuberculosis in those who were in the R form, rather than S form, positively reacted. The use of FV identified 2 groups of hybridomas obtained after immunization of albino mice with the capsule-free variant of a plague microbe. Some hybridomas reacted only with plague bacteria while others did with two above pathogens. The authors substantiate the expediency of using FV, its components, and obtained monoclonal plague pathogen antibodies to improve antiplague diagnosticums with an activity spectrum that exceeds that of the existing commercial F1 antigen-based diagnosticums. They also discuss the lines of further studies.
对通过在梯度表面活性剂中从鼠疫杆菌无荚膜疫苗株EV76的活细菌和丙酮干燥细菌中提取而分离得到的实验性抗原复合物进行了研究，该疫苗株已丧失合成诊断性种特异性荚膜抗原F1的能力。在表面活性剂浓度为1.28%的第五阶段提取及进一步纯化后，获得了抗原复合物组分V（FV）。发现热稳定的FV主要由蛋白质组成。该组分中分子量约为43 kD的蛋白质占主导。后者对白化小鼠无毒且具有抗原性。它与市售抗鼠疫血清抗体形成沉淀。用FV对抗鞣制绵羊红细胞进行抗原致敏后得到一种诊断试剂，该试剂与抗鼠疫血清特异性反应，而不与抗肠道杆菌的异源血清反应。用FV免疫的血清在JDJFR中与所有鼠疫病原菌菌株特异性反应，无论其培养温度如何，包括“无组分”菌株，该菌株不与F1诊断试剂相互作用。用无荚膜鼠疫微生物株免疫的动物血清仅与FV - 红细胞诊断试剂反应，而不与F1抗原致敏的红细胞相互作用。在ABNR中用130株典型和非典型鼠疫微生物菌株以及133株肠杆菌科不同种的异源细菌对红细胞FV诊断试剂进行了测试。FV诊断试剂识别出所有鼠疫杆菌变体，而F1诊断试剂仅揭示其荚膜变体。在异源细菌中，处于R型而非S型的密切相关的假结核病原菌的一些菌株呈阳性反应。使用FV鉴定了用鼠疫杆菌无荚膜变体免疫白化小鼠后获得的2组杂交瘤。一些杂交瘤仅与鼠疫杆菌反应，而其他杂交瘤则与上述两种病原菌反应。作者证实了使用FV及其组分以及获得的单克隆鼠疫病原菌抗体来改进抗鼠疫诊断试剂的合理性，其活性谱超过了现有的基于商业F1抗原的诊断试剂。他们还讨论了进一步研究的方向。
