Sacchi C T, de Lemos A P, Whitney A M, Melles C E, Solari C A, Frasch C E, Mayer L W
Bacteriology Division, Adolfo Lutz Institute, São Paulo, Brazil.
Rev Inst Med Trop Sao Paulo. 1998 Mar-Apr;40(2):113-7. doi: 10.1590/s0036-46651998000200009.
In the present study we examine the potential use of oligonucleotide probes to characterize Neisseria meningitidis serotypes without the use of monoclonal antibodies (MAbs). Antigenic diversity on PorB protein forms the bases of serotyping method. However, the current panel of MAbs underestimated, by at least 50% the PorB variability, presumably because reagents for several PorB variable regions (VRs) are lacking, or because a number of VR variants are not recognized by serotype-defining MAbs. We analyzed the use of oligonucleotide probes to characterize serotype 10 and serotype 19 of N. meningitidis. The porB gene sequence for the prototype strain of serotype 10 was determined, aligned with 7 other porB sequences from different serotypes, and analysis of individual VRs were performed. The results of DNA probes 21U (VR1-A) and 615U (VR3-B) used against 72 N. meningitidis strains confirm that VR1 type A and VR3 type B encode epitopes for serotype-defined MAbs 19 and 10, respectively. The use of probes for characterizing serotypes possible can type 100% of the PorB VR diversity. It is a simple and rapid method specially useful for analysis of large number of samples.
在本研究中，我们探讨了寡核苷酸探针在不使用单克隆抗体（MAb）的情况下对脑膜炎奈瑟菌血清型进行鉴定的潜在用途。PorB蛋白上的抗原多样性构成了血清分型方法的基础。然而，目前的单克隆抗体组至少低估了PorB变异性的50%，推测原因是缺乏针对几个PorB可变区（VR）的试剂，或者是一些VR变体未被血清型定义单克隆抗体识别。我们分析了寡核苷酸探针用于鉴定脑膜炎奈瑟菌血清型10和血清型19的用途。测定了血清型10原型菌株的porB基因序列，将其与来自不同血清型的其他7个porB序列进行比对，并对各个VR进行了分析。针对72株脑膜炎奈瑟菌菌株使用DNA探针21U（VR1-A）和615U（VR3-B）的结果证实，VR1 A型和VR3 B型分别编码血清型定义单克隆抗体19和10的表位。使用探针鉴定血清型有可能对100%的PorB VR多样性进行分型。这是一种简单快速的方法，特别适用于大量样本的分析。
