Improved allele-specific PCR assays for detection of clarithromycin and fluoroquinolone resistant of Helicobacter pylori in gastric biopsies: identification of N87I mutation in GyrA.
Improved allele-specific PCR assays for detection of clarithromycin and fluoroquinolone resistant of Helicobacter pylori in gastric biopsies: identification of N87I mutation in GyrA.
复制标题
DOI：
10.1016/j.diagmicrobio.2014.12.003
复制
发表时间：
2015-04
影响因子：
2.9
通讯作者：
Graham DY
中科院分区：
文献类型：
--
作者：
Trespalacios AA;Rimbara E;Otero W;Reddy R;Graham DY
分子检测可以通过胃活检快速检测幽门螺杆菌的敏感性。用等位基因特异性聚合酶链式反应(ASP-PCR)检测哥伦比亚患者胃组织中Hp的23S rRNA和gyrA突变，并经23S rRNA和gyrA基因测序证实。用琼脂稀释法测定其敏感性和特异性，比较其敏感性和特异性。样本包括107例幽门螺杆菌感染的胃活检组织和20例幽门螺杆菌阴性的活检组织。ASP-PCR检测23S rRNA基因的敏感性和特异性均为100%。2007年Nishizawa等人发表的GyrA基因ASP-PCR的敏感性和特异性分别为52%和92.7%，敏感性较低是由于我们的样本中存在N87I突变，而本试验没有检测到N87I突变。在本研究中，我们设计了新的引物来检测GyrA中的突变N87I。用原有的引物和新的引物进行ASP-PCR扩增。经分子检测，灵敏度提高到100%。总之，ASP-PCR为预测胃活检组织对克拉霉素和左氧氟沙星的耐药性提供了一种特异和快速的手段。
Molecular testing can rapidly detect Helicobacter pylori susceptibility using gastric biopsies. Allele-specific polymerase chain reaction (ASP-PCR) was used to identify H. pylori 23S rRNA and gyrA mutation using gastric biopsies from Colombian patients and confirmed by PCR and sequencing of the 23S rRNA and gyrA genes. The sensitivity and specificity of ASP-PCR were compared with susceptibilities measured by agar dilution. Samples included gastric biopsies from 107 biopsies with H. pylori infections and 20 H. pylori negative. The sensitivity and specificity of ASP-PCR for the 23S rRNA gene were both 100%. The sensitivity and specificity of ASP-PCR for the gyrA gene, published in 2007 by Nishizawa et al., were 52% and 92.7%, respectively; the lower sensitivity was due to the presence of mutation N87I in our samples, which were not detected by the test. In this study, we designed new primers to detect the mutation N87I in GyrA. The ASP-PCR was performed with the original primers plus the new primers. The molecular test with the new primers improved the sensitivity to 100%. In conclusion, ASP-PCR provides a specific and rapid means of predicting resistance to clarithromycin and levofloxacin in gastric biopsies.
登录
查看更多内容
DOI：
10.1016/j.ijantimicag.2006.11.007
发表时间：
2007-04-01
影响因子：
10.8
作者：
Cattoir, Vincent;Nectoux, Juliette;Cambau, Emmanuelle
通讯作者：
Cambau, Emmanuelle
DOI：
10.1111/j.1440-1746.2007.04919.x
发表时间：
2007-11-01
影响因子：
4.1
作者：
Furuta, Takahisa;Soya, Yoshihiro;Hishida, Akira
通讯作者：
Hishida, Akira
影响因子：
3
作者：
De Francesco, Vincenzo;Zullo, Angelo;Vaira, Dino
通讯作者：
Vaira, Dino
影响因子：
4.4
作者：
Miyachi, Hideyuki;Miki, Ikuya;Kasuga, Masato
通讯作者：
Kasuga, Masato
影响因子：
2.4
作者：
Lehours, Philippe;Siffre, Elodie;Megraud, Francis
通讯作者：
Megraud, Francis