Hinrikson H P, Dutly F, Altwegg M
Department of Medical Microbiology, University of Zürich, CH-8028 Z ürich, Switzerland.
J Clin Microbiol. 1999 Jan;37(1):152-6. doi: 10.1128/JCM.37.1.152-156.1999.
The current genetic strategies used to identify Tropheryma whippelii, the putative agent of Whipple's disease, are based on PCR-mediated amplification of a part of its 16S rRNA gene (16S rDNA). Because there is very little intraspecies variation in these molecules, they are not suitable as targets for epidemiologic investigations. However, the intergenic spacer region between the 16S and 23S rDNAs is usually much more variable and has repeatedly been used for epidemiologic purposes. We have therefore amplified the spacer region of T. whippelii directly from clinical specimens from nine independent Swiss patients with Whipple's disease by PCR with primers complementary to the 3' and 5' ends of the 16S and 23S rDNAs, respectively. The amplicons were directly sequenced and the sequences were compared to the T. whippelii reference sequence in GenBank/EMBL (accession no. X99636). Complete sequence homogeneity was found between the samples from our nine patients; the spacer sequence was also identical to the reference sequence. However, the sequences corresponding to the 3' and 5' ends of the 16S and the 23S rDNAs of T. whippelii, respectively, differed from the respective sequences in GenBank/EMBL. The same sequence found in our patients was then found in a sample from the German patient from which the published sequence had been derived. We conclude that the 16S-23S rDNA spacer region seems to be very conserved in T. whippelii and that the respective reference entry in public databases should be revised.
目前用于鉴定惠普尔病假定病原体惠普尔嗜组织菌的基因策略,是基于聚合酶链反应(PCR)介导的对其16S核糖体RNA基因(16S rDNA)一部分的扩增。由于这些分子在种内变异非常小,它们不适合作为流行病学调查的靶点。然而,16S和23S rDNA之间的基因间隔区通常变异更大,并且已多次用于流行病学目的。因此,我们通过PCR,分别使用与16S和23S rDNA的3'端和5'端互补的引物,直接从9名瑞士惠普尔病独立患者的临床标本中扩增了惠普尔嗜组织菌的间隔区。对扩增产物进行直接测序,并将序列与GenBank/EMBL中惠普尔嗜组织菌的参考序列(登录号X99636)进行比较。在我们9名患者的样本之间发现了完全的序列同源性;间隔区序列也与参考序列相同。然而,分别对应于惠普尔嗜组织菌16S和23S rDNA的3'端和5'端的序列,与GenBank/EMBL中的相应序列不同。然后在来自德国患者的一个样本中发现了与我们患者中相同的序列,已发表的序列即源自该德国患者的样本。我们得出结论,16S - 23S rDNA间隔区在惠普尔嗜组织菌中似乎非常保守,公共数据库中的相应参考条目应予以修订。