以Real-time PCR方法检测耐药幽门螺杆菌中HefABC基因的表达

HefABC Expression of Antibiotic Resistant Helicobacter pylori Determined by Real-time PCR

为了探讨耐药幽门螺杆菌(Helicobacter. pylori)主动外排系统hefA、hefB、hefC基因表达与菌株克拉霉素耐药、阿莫西林耐药以及多重耐药的关系,随机选择H. pylori克拉霉素耐药株、阿莫西林耐药株、敏感株、多重耐药株各10株,利用Real-time PCR方法检测HefABC系统的表达,以成组t检验法对敏感株与各耐药株之间mRNA表达水平进行比较。结果显示,hefA在敏感组、克拉霉素耐药组、阿莫西林耐药组、多重耐药组中的mRNA表达量以2-ΔΔCt表示,分别为0.903 9±0.154 07、1.482 2±0.378 5、1.278 0±0.206 2、2.255 7±0.289 2,克拉霉素耐药组与敏感组比较:t=4.489,p<0.05;阿莫西林耐药组与敏感组比较:t=4.631,p<0.05;多重耐药组与敏感组比较:t=13.11,p<0.05。hefB在敏感组、克拉霉素耐药组、阿莫西林耐药组、多重耐药组中的m RNA表达量以2-ΔΔCt表示,分别为0.075 9±0.015 7、0.636 1±0.162 3、0.906 4±0.127 6、1.149 2±0.305 8,克拉霉素耐药组与敏感组比较:t=10.86,p<0.05;阿莫西林耐药组与敏感组比较:t=20.42,p<0.05;多重耐药组与敏感组比较:t=11.09,p<0.05。hefC在敏感组、克拉霉素耐药组、阿莫西林耐药组、多重耐药组中的m RNA表达量以2-ΔΔCt表示,分别为0.128 2±0.054 5、1.319 4±0.272 4、0.683 0±0.185 0、3.480 2±0.932 6;克拉霉素耐药组与敏感组比较:t=13.56,p<0.05;阿莫西林耐药组与敏感组比较:t=9.096,p<0.05;多重耐药组与敏感组比较:t=11.35,p<0.05。研究结果证实,H. pylori中外排系统hefA、hefB、hefC的高表达与菌株对克拉霉素耐药、阿莫西林耐药及多重耐药均有关。

This study aimed to explore the role of hefA,hefB and hefC gene expression of H.pylori in amoxicillin resistance,clarithromycin resistance and multiple drug resistance.10 strains of amoxicillin-resistant strains,10 strains of clarithromycin resistant strains,10 strains of susceptible strains and 10 multidrug resistant strains were randomly selected to detect the expression of HefABC system by Real-time PCR method.The m RNA expression levels between susceptible and resistant strains were compared,and statistical analysis was carried out by t test.Expression levels of hefA gene in each group were as follows:susceptible group(0.903 9±0.1540 7),clarithromycin-resistant group(1.482 2±0.378 5),amoxicillin-resistant group(1.278 0±0.206 2)and multidrug-resistant group(2.255 7±0.289 2).The expression of resistant strains of clarithromycin was higher than the susceptible strains(t=4.489,p<0.05).The expression of resistant strains of amoxicillin was higher than the susceptible strains(t=4.631,p<0.05).The expression of resistant strains of multidrug-resistant was significantly higher than the susceptible strains(t=13.11,p<0.05).Expression levels of hefB in each group were as follows:susceptible group(0.075 9±0.015 7),clarithromycin-resistant group(0.636 1±0.162 3),amoxicillin-resistant group(0.906 4±0.127 6)and multidrug-resistant group(1.149 2±0.305 8).The expression of resistant strains of clarithromycin was higher than the susceptible strains(t=10.86,p<0.05).The expression of resistant strains of amoxicillin was higher than the susceptible strains(t=20.42,p<0.05).The expression of resistant strains of multidrug-resistant was significantly higher than the susceptible strains(t=11.09,p<0.05)).Expression levels of hefC in each group were as follows:susceptible group(0.128 2±0.054 5),clarithromycin-resistant group(1.319 4±0.272 4),amoxicillin-resistant group(0.683 0±0.185 0)and multidrug-resistant group(3.480 2±0.932 6).The expression of resistant strains of clarithromycin was higher than the susceptible strains(t=13.56,p<0.05)).The expression of resistant strains of amoxicillin was higher than the susceptible strains(t=9.096,p<0.05)).The expression of resistant strains of multidrug-resistant was significantly higher than the susce ptible strains(t=11.35,p<0.05)).The high expression of hefA,hefB,hefC in the efflux system is related to H.pylori resistance to clarithromycin,amoxicillin resistance and multiple drug resistance.