Umegaki N, Shimoyama T, Nishiya D, Suto T, Fukuda S, Munakata A
First Department of Internal Medicine, Hirosaki University School of Medicine, Japan.
J Gastroenterol Hepatol. 2000 Aug;15(8):906-9. doi: 10.1046/j.1440-1746.2000.02072.x.
Resistance of Helicobacter pylori to clarithromycin is mostly due to the point mutations in the 23S rRNA. In Japan, however, the frequency of these mutations has not been fully investigated. Furthermore, no study has used gastric biopsy specimens to detect these point mutations.
The frequency of primary clarithromycin-resistant H. pylori was examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Eighty-two strains (42 isolated from patients with gastric cancer and 40 isolated from patients with chronic gastritis) were examined. Two biopsy specimens obtained from patients in whom eradication therapy including clarithromycin had failed were also studied.
Either A2143G or A2144G point mutation was detected in 90% of clarithromycin-resistant H. pylori strains. Eight out of 82 strains (9.8%) had either A2143G or A2144G point mutation. Only one out of 42 strains in patients with gastric cancer had A2143G mutation, whereas five strains had A2144G and two had A2143G mutations in 40 strains isolated from control subjects. The proportion was significantly lower in patients with early gastric cancer (P < 0.05). This PCR-RFLP was also applicable for DNA samples extracted from biopsy specimens and infection of clarithromycin-resistant H. pylori was observed.
The results suggest that the point mutation in the 23S rRNA gene is commonly seen in clarithromycin-resistant H. pylori and it contributes to the treatment failure in Japan. The PCR-RFLP system is a sensitive method by which to diagnose H. pylori infection as well as a simple method for detecting clarithromycin resistance without bacterial culture.
幽门螺杆菌对克拉霉素的耐药性主要归因于23S rRNA中的点突变。然而,在日本,这些突变的频率尚未得到充分研究。此外,尚无研究使用胃活检标本检测这些点突变。
采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)检测原发性克拉霉素耐药幽门螺杆菌的频率。检测了82株菌株(42株从胃癌患者中分离,40株从慢性胃炎患者中分离)。还研究了从接受包括克拉霉素在内的根除治疗失败的患者中获取的两份活检标本。
在90%的克拉霉素耐药幽门螺杆菌菌株中检测到A2143G或A2144G点突变。82株菌株中有8株(9.8%)发生了A2143G或A2144G点突变。胃癌患者的42株菌株中只有1株发生了A2143G突变,而在从对照受试者中分离的40株菌株中,有5株发生了A2144G突变,2株发生了A2143G突变。早期胃癌患者中的比例显著较低(P<0.05)。这种PCR-RFLP也适用于从活检标本中提取的DNA样本,并且观察到了克拉霉素耐药幽门螺杆菌的感染。
结果表明,23S rRNA基因中的点突变在克拉霉素耐药幽门螺杆菌中常见,并且在日本导致了治疗失败。PCR-RFLP系统是一种诊断幽门螺杆菌感染的敏感方法,也是一种无需细菌培养即可检测克拉霉素耐药性的简单方法。