幽门螺杆菌临床分离株耐药特性分析及嗜酸乳杆菌对耐药幽门螺杆菌的抑制作用

目的 对84株临床分离幽门螺杆菌 (Helicobacter pylori,Hp)菌株进行耐药特性分析,并同时观察抗Hp嗜酸乳杆菌(Lactobacillus acidophilus,La)4和La6对不同抗生素耐药Hp的抑制效应.方法 从84例不同胃病(慢性胃炎25例,胃溃疡24例,十二指肠溃疡19例,胃癌16例)患者胃黏膜中分离培养Hp菌株,采用E-test法测定甲硝唑、克拉霉素和阿莫西林3种常用抗生素对临床Hp菌株的最小抑菌浓度(MIC值),了解临床Hp菌株对3种抗生素的耐药状况.以标准La作对照,将抗Hp La4和La6上清液加入含不同Hp菌株培养板的孔中,固体培养72 h,记录抑菌环;抗Hp La菌液加入含不同Hp 菌株的培养液中,液体培养,于不同时间点(4、8、12、24和48 h)分别取培养液,计算菌落形成单位和测定尿素酶活性.结果 84株临床分离Hp菌株中对甲硝唑、克拉霉素和阿莫西林的耐药率分别为67.9%、17.9%和1.2%.其中11株为混合耐药,包括 10株甲硝唑和克拉霉素混合耐药和1株甲硝唑和阿莫西林混合耐药.在固体培养条件下抗Hp La4和La6上清液对抗生素耐药和非耐药Hp菌株均产生明显的抑菌效应.液体培养条件下抗Hp La4和La6菌液能抑制抗生素耐药和非耐药Hp菌株的增殖,其拮抗Hp的作用明显强于标准La菌株(P<0.01).抗Hp La4和La6在混合培养4 h就能抑制抗生素耐药Hp菌株尿素酶活性,随着培养时间的延长Hp尿素酶活性逐渐降低,其抑制作用明显强于标准La组(P＜0.05).结论 84株临床分离Hp菌株中甲硝唑耐药株为常见,其次为克拉霉素耐药和甲硝唑克拉霉素混合耐药株.抗Hp La4和La6在体外对抗生素耐药和非耐药Hp菌株均具有明显的抑制作用.

Abstract：

Objective To analyze the drug resistant characteristics of 84 clinical isolated Helicobacter pylori (Hp) strains, and to observe the inhibitory effects of anti-Hp Lactobacillus acidophilus (La)4 and La6 on different antibiotic-resistant Hp strains. Methods Hp strains were isolated and cultured from gastric mucosa of 84 different gastropathy patients (20 patients with chronic gastritis, 24 with gastric ulcer, 19 with duodenal ulcer and 16 with gastric cancer). The minimum inhibitory concentration (MIC) of metronidazole, clarithromycin and amoxicillin were tested by E-test in order to determine the resistance of these three antibiotics in clinical isolated Hp strains. With standard La as control, the supernatant of anti-Hp La4 and La6 was added into Hp strains culture wells. Hp strains were cultured in solid media for 72 hours, and then inhibition ring were recorded. Anti-Hp Lactobacillus acidophilus liquid was also added to culture medium of different Hp strains, which were in liquid culture, culture medium were taken at different time points (4,8,12,24,48 hrs) to calculate bacteria colony number and test urease activity. Results In 84 clinical isolated Hp strains, the resistant rates of metronidazole, clarithromycin and amoxicillin resistance rates were 67.9%, 17.9% and 1.2% respectively. Of those 11 strains were mixed drug resistance, which included 10 strains of metronidazole and clarithromycin mixed drug resistance, and one of metronidazole and amoxicillin mixed drug resistance. In solid culture conditions, supernatant of anti-Hp Lactobacillus acidophilus La4 and La6 had obvious inhibitory effect on antibiotic-resistant and non-resistant Hp strains. In liquid culture conditions, anti-Hp Lactobacillu acidophilus La4 and La6 bacterium liquid could inhibit the proliferation of antibiotic-resistant and non-resistant Hp strains, the antagonistic role was significantly stronger than the standard Lactobacillus acidophilus strains (P<0.05). The urease activity of antibiotic-resistant Hp strains was inhibited since mixed cultured with anti-Hp Lactobacillu acidophilus La4 and La6 for 4 hours, the urease activity gradually decreased as culture time extended, and the inhibitory role was significantly stronger than the standard Lactobacillus acidophilus strains (P<0.05). Conclusions In 84 Hp strains, most were metronidazole resistant strains, followed by clarithromycin resistant strains, metronidazole and clarithromycin mixed resistance strains. In vitro, anti-Hp Lactobacillu acidophilus La4 and La6 had obvious inhibitory effects on antibiotic-resistant and non-resistant Hp strains.

The analysis of drug resistance characteristics in clinical isolated Helicobacter pylori strains from different gastropathy patients and the inhibitory effect of Lactobacillus acidophilus on drug-resistant Helicobacter pylori

Objective To analyze the drug resistant characteristics of 84 clinical isolated Helicobacter pylori (Hp) strains, and to observe the inhibitory effects of anti-Hp Lactobacillus acidophilus (La)4 and La6 on different antibiotic-resistant Hp strains. Methods Hp strains were isolated and cultured from gastric mucosa of 84 different gastropathy patients (20 patients with chronic gastritis, 24 with gastric ulcer, 19 with duodenal ulcer and 16 with gastric cancer). The minimum inhibitory concentration (MIC) of metronidazole, clarithromycin and amoxicillin were tested by E-test in order to determine the resistance of these three antibiotics in clinical isolated Hp strains. With standard La as control, the supernatant of anti-Hp La4 and La6 was added into Hp strains culture wells. Hp strains were cultured in solid media for 72 hours, and then inhibition ring were recorded. Anti-Hp Lactobacillus acidophilus liquid was also added to culture medium of different Hp strains, which were in liquid culture, culture medium were taken at different time points (4,8,12,24,48 hrs) to calculate bacteria colony number and test urease activity. Results In 84 clinical isolated Hp strains, the resistant rates of metronidazole, clarithromycin and amoxicillin resistance rates were 67.9%, 17.9% and 1.2% respectively. Of those 11 strains were mixed drug resistance, which included 10 strains of metronidazole and clarithromycin mixed drug resistance, and one of metronidazole and amoxicillin mixed drug resistance. In solid culture conditions, supernatant of anti-Hp Lactobacillus acidophilus La4 and La6 had obvious inhibitory effect on antibiotic-resistant and non-resistant Hp strains. In liquid culture conditions, anti-Hp Lactobacillu acidophilus La4 and La6 bacterium liquid could inhibit the proliferation of antibiotic-resistant and non-resistant Hp strains, the antagonistic role was significantly stronger than the standard Lactobacillus acidophilus strains (P<0.05). The urease activity of antibiotic-resistant Hp strains was inhibited since mixed cultured with anti-Hp Lactobacillu acidophilus La4 and La6 for 4 hours, the urease activity gradually decreased as culture time extended, and the inhibitory role was significantly stronger than the standard Lactobacillus acidophilus strains (P<0.05). Conclusions In 84 Hp strains, most were metronidazole resistant strains, followed by clarithromycin resistant strains, metronidazole and clarithromycin mixed resistance strains. In vitro, anti-Hp Lactobacillu acidophilus La4 and La6 had obvious inhibitory effects on antibiotic-resistant and non-resistant Hp strains.