Graham T, Golsteyn-Thomas E J, Gannon V P, Thomas J E
Agriculture and Agri-Food Canada, Lethbridge, AB, Canada.
Can J Microbiol. 1996 Nov;42(11):1155-62. doi: 10.1139/m96-147.
In this study, the 16S/23S rRNA intergenic spacer (IGS) regions of six Listeria species were examined. DNA bands of 590 and 340 bp were observed following polymerase chain reaction (PCR) amplification of DNA from Listeria monocytogenes, Listeria innocua, Listeria seeligeri, Listeria welshimeri, and Listeria ivanovii strains with generic rRNA IGS oligonucleotide primers. For strains of Listeria grayi subspp. grayi and murrayi, DNA band sizes of 550 and 340 bp were observed with this primer pair. DNA bands of these sizes were not observed for other Gram-negative or- positive bacteria in this PCR assay. Four RsaI digestion profiles were noted for the Listeria PCR products. Listeria monocytogenes strains had one profile; L. innocua strains had a second; L seeligeri, L. welshimeri, and L ivanovii strains had a third; and L. grayi strains had a fourth. The small and large 16S/23S rRNA IGSs of L. monocytogenes ATCC 15313 were identical in the first 58 5' and the last 169 3' nucleotides. However, the large rRNA IGS contained a central 267-bp region with tRNA(Ile) and tRNA(Ala) genes. Large rRNA 16S/23S IGS nucleotide sequence data has not been previously reported. This data was used to develop novel Listeria genus-specific and L.monocytogenes species-specific PCR assays.
在本研究中，检测了六种李斯特菌属细菌的16S/23S核糖体RNA基因间隔区（IGS）。用通用核糖体RNA IGS寡核苷酸引物对单核细胞增生李斯特菌、无害李斯特菌、斯氏李斯特菌、威尔氏李斯特菌和伊氏李斯特菌菌株的DNA进行聚合酶链反应（PCR）扩增后，观察到590和340 bp的DNA条带。对于格氏李斯特菌格氏亚种和默里亚种的菌株，用该引物对观察到的DNA条带大小为550和340 bp。在该PCR检测中，未观察到其他革兰氏阴性或阳性细菌有这些大小的DNA条带。注意到李斯特菌PCR产物有四种RsaI酶切图谱。单核细胞增生李斯特菌菌株有一个图谱；无害李斯特菌菌株有第二个图谱；斯氏李斯特菌、威尔氏李斯特菌和伊氏李斯特菌菌株有第三个图谱；格氏李斯特菌菌株有第四个图谱。单核细胞增生李斯特菌ATCC 15313的小16S/23S rRNA IGS和大16S/23S rRNA IGS在前58个5'核苷酸和最后169个3'核苷酸处是相同的。然而，大rRNA IGS包含一个带有tRNA（Ile）和tRNA（Ala）基因的267 bp中央区域。大rRNA 16S/23S IGS核苷酸序列数据此前尚未见报道。该数据用于开发新的李斯特菌属特异性和单核细胞增生李斯特菌种特异性PCR检测方法。
