101例胃黏膜幽门螺杆菌感染耐药基因突变分析

Gene mutation and drug resistance analysis for 101 cases of gastric Helicobacter pylori infection

目的:比较荧光定量PCR方法与免疫组织化学法检测胃黏膜活检样本中幽门螺杆菌(Helicobacter pylori, Hp)感染率,评估荧光定量PCR方法检测Hp对克拉霉素和左氧氟沙星耐药性的可行性。方法:分别通过荧光定量PCR方法与免疫组织化学法检测101例胃黏膜Hp感染情况,并比较两种方法检测Hp的阳性率,然后通过荧光定量PCR法检测Hp阳性样本中23S rRNA和GYRA的基因突变情况,再用Sanger测序法分析23S rRNA基因与GYRA基因突变类型。结果:101例胃黏膜样本中PCR法检测出Hp阳性70例(69.31%),Hp阴性31例(30.69%);免疫组织化学法检测出Hp阳性68例(67.33%),Hp阴性33例(32.67%);两种方法检测Hp阳性率比较差异无统计学意义(P>0.05)。70例荧光定量PCR法检测Hp阳性样本中克拉霉素耐药30例(42.25%),左氧氟沙星耐药15例(21.13%),其中有8例(11.27%)对克拉霉素与左氧氟沙星双重耐药,克拉霉素与左氧氟沙星敏感型33例(47.14%)。结论:荧光定量PCR方法与免疫组织化学法检测Hp均具备较好的阳性检出率,荧光定量PCR法不仅可以检测Hp感染还能进行抗Hp相关抗生素的耐药性检测,具有快速、准确的特点,对指导个体化治疗具有临床意义。

Objective:To compare the infection rate of Helicobacter pylori(Hp) in gastric mucosal biopsy samples by fluorescence quantitative PCR and immunohistochemistry,and to evaluate the feasibility of fluorescence quantitative PCR in detecting the resistance of Hp to clarithromycin and levofloxacin.Methods:A total of 101 cases of Hp infection in gastric mucosa were detected by fluorescence quantitative PCR and immunohistochemistry,and the infection rates of the 2 methods were compared.Thhen,23 S rRNA and gyrA gene mutations in Hp positive samples were detected by fluorescence quantitative PCR,and the mutation types of 23 S rRNA gene and gyrA gene were analyzed by Sanger sequencing.Results:Among 101 gastric mucosa samples,70 cases(69.31%) were Hp positive and 31 cases(30.69%) were Hp negative by PCR;68 cases(67.33%) were Hp positive and 33 cases(32.67%) were Hp negative by immunohistochemistry.There was no significant difference in the positive rate of Hp between the 2 methods(P>0.05);Among 70 Hp positive samples detected by fluorescence quantitative PCR,30 cases were clarithromycin resistant(42.25%),15 cases were levofloxacin resistant(21.13%),of which 8 cases(11.27%) were dual resistant to both clarithromycin and levofloxacin.Besides,33 cases were sensitive to clarithromycin and levofloxacin(47.14%).Conclusion:Both fluorescence quantitative PCR and immunohistochemistry have a good positive rate in the detection of Hp.Fluorescence quantitative PCR can not only detect Hp infection,but also reveal the drug resistance of anti Hp related antibiotics.PCR shows good rapidity and accuracy,with great clinical significance in guiding individualized treatment.