Tazumi A, Negoro M, Tomiyama Y, Misawa N, Itoh K, Moore J E, Millar B C, Matsuda M
Laboratory of Molecular Biology, School of Environmental Health Sciences, Azabu University, Sagamihara, Japan.
Br J Biomed Sci. 2011;68(1):19-22. doi: 10.1080/09674845.2011.11732836.
Polymerase chain reaction (PCR) amplification was performed on 20 isolates of five Campylobacter species using a degenerate primer pair designed in silico to generate a product of the luxS gene or its homologue from Campylobacter organisms. Although the primer pair successfully amplified products of approximately 500 base pairs (bp) with the eight isolates of C. jejuni and C. coli and some of C. upsaliensis and C. fetus, it failed to amplify fragments with all four isolates of C. lari (two urease-negative C. lari; two urease-positive thermophilic campylobacters). When Southern blot hybridisation analysis was carried using the mixed luxS gene fragments prepared from the C. jejuni, C. coli, C. upsaliensis and C. fetus strains as a probe, all C. jejuni, C. coli, C. upsaliensis and C. fetus isolates gave positive signals, but no positive signal was detected with any C. lari isolate. These results clearly indicate that C. jejuni, C. coli, C. upsaliensis and C. fetus carry the luxS gene or its homologue. However, no luxS gene or its homologue was identified to occur in the C. lari genome. Although autoinducer-2 assays were positive in C. jejuni, C. coli, C. upsaliensis and C. fetus isolates, it was negative with all the C. lari isolates examined. In addition, a biofilm formation assay demonstrated that biofilm formation in the C. lari species does not appear to correlate with the occurrence of the luxS gene because biofilm formation occurred among some isolates of C. lari.
使用在计算机上设计的简并引物对,对5种弯曲杆菌属细菌的20个分离株进行聚合酶链反应(PCR)扩增,以从弯曲杆菌属生物体中产生luxS基因或其同源物的产物。尽管该引物对成功地用8株空肠弯曲菌、结肠弯曲菌以及一些乌普萨拉弯曲菌和胎儿弯曲菌分离株扩增出了约500个碱基对(bp)的产物,但未能用所有4株拉氏弯曲菌分离株(2株脲酶阴性拉氏弯曲菌;2株脲酶阳性嗜热弯曲杆菌)扩增出片段。当使用从空肠弯曲菌、结肠弯曲菌、乌普萨拉弯曲菌和胎儿弯曲菌菌株制备的混合luxS基因片段作为探针进行Southern印迹杂交分析时,所有空肠弯曲菌、结肠弯曲菌、乌普萨拉弯曲菌和胎儿弯曲菌分离株均给出阳性信号,但未检测到任何拉氏弯曲菌分离株的阳性信号。这些结果清楚地表明,空肠弯曲菌、结肠弯曲菌、乌普萨拉弯曲菌和胎儿弯曲菌携带luxS基因或其同源物。然而,未在拉氏弯曲菌基因组中鉴定出luxS基因或其同源物。尽管自诱导物-2检测在空肠弯曲菌、结肠弯曲菌、乌普萨拉弯曲菌和胎儿弯曲菌分离株中呈阳性,但在所检测的所有拉氏弯曲菌分离株中均为阴性。此外,生物膜形成试验表明,拉氏弯曲菌属中的生物膜形成似乎与luxS基因的存在无关,因为在一些拉氏弯曲菌分离株中发生了生物膜形成。