John G H, Carlson J O, Kimberling C V, Ellis R P
Department of Microbiology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins 80523.
J Clin Microbiol. 1990 Nov;28(11):2456-61. doi: 10.1128/jcm.28.11.2456-2461.1990.
Serogrouping of Bacteroides nodosus is based on antigenic differences in fimbriae of the different New Zealand prototype strains. Because of the time needed to isolate and grow pure cultures of B. nodosus and the difficulty in distinguishing between different serogroups because of cross-agglutination, a new DNA-based diagnostic approach based on the fimbrial gene sequence of B. nodosus was developed. Published nucleotide sequences of the fimbrial genes for serogroups A, G, D, and H showed conservation at the 5' end, coding for the N terminus, and variability at the 3' end, coding for the C terminus. The polymerase chain reaction was used to amplify both the constant and variable regions of the fimbrial genes. Constant-region oligonucleotide primers were used to amplify a 100-base-pair fragment from the constant regions of the fimbrial genes of 10 New Zealand serogroups. Serogroup-specific oligonucleotide primers for serogroups A and H allowed amplification of a 282-base-pair fragment from serogroup A and a 363-base-pair fragment from serogroup H. Thus, amplification of the constant and variable regions of the fimbrial gene allows rapid detection and grouping of B. nodosus.
结节拟杆菌的血清分型基于不同新西兰原型菌株菌毛的抗原差异。由于分离和培养结节拟杆菌纯培养物所需的时间，以及因交叉凝集难以区分不同血清型，因此开发了一种基于结节拟杆菌菌毛基因序列的新型DNA诊断方法。已发表的A、G、D和H血清型菌毛基因的核苷酸序列显示，在编码N端的5'端具有保守性，而在编码C端的3'端具有变异性。聚合酶链反应用于扩增菌毛基因的恒定区和可变区。使用恒定区寡核苷酸引物从10个新西兰血清型的菌毛基因恒定区扩增出一个100个碱基对的片段。A和H血清型的血清型特异性寡核苷酸引物可分别从A血清型扩增出一个282个碱基对的片段，从H血清型扩增出一个363个碱基对的片段。因此，扩增菌毛基因的恒定区和可变区可实现结节拟杆菌的快速检测和分型。
