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快速尿素酶试验样本用于幽门螺杆菌克拉霉素耐药性分子分析的效用

Usefulness of rapid urease test samples for molecular analysis of clarithromycin resistance in Helicobacter pylori.

作者信息

Baroni María R, Bucci Pamela, Giani Rita N, Giusti Antonela, Tedeschi Fabian A, Salvatierra Emiliano, Barbaglia Yanina, Jimenez Félix, Zalazar Fabian E

机构信息

Laboratorio de Práctica Profesional de Bioquímica, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral/Hospital "Dr. José María Cullen", Santa Fe, Argentina.

Laboratorio de Práctica Profesional de Bioquímica, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral/Hospital "Dr. José María Cullen", Santa Fe, Argentina.

出版信息

Rev Argent Microbiol. 2018 Oct-Dec;50(4):359-364. doi: 10.1016/j.ram.2017.11.005. Epub 2018 Mar 27.

Abstract

Helicobacter pylori is a gastric pathogen that is widely recognized as a causative agent of gastric disease. Its eradication is variable, mainly due to increased resistance to clarithromycin. Our objective was: to evaluate (i) if the biopsy specimen used for the rapid urease test is a useful sample to detect resistance to clarithromycin by PCR-RFLP and (ii) the distribution of A2142G and A2143G point mutations in the 23S rRNA gene, in relation to virulence factors in our region. Gastric specimens were collected from adult dyspeptic patients (n=141) and H. pylori was investigated by the rapid urease test, histopathological analysis and PCR for the hsp60 gene. Clarithromycin resistance was detected by PCR-RFLP in 62 H. pylori (+) paired biopsy specimens submitted to molecular analysis and the rapid urease test. H. pylori virulence factors were analyzed by multiplex PCR using specific primers for the cagA, vacA and babA2 genes. Thirteen out of 62 strains (20.9%) were resistant to clarithromycin: 6/13 (46.2%) harbored the A2143G mutation whereas 7/13 (53.8%) carried the A2142G point mutation. vacA m1s1 was the most frequent genotype among the resistant strains. In conclusion, the biopsy specimens used for the rapid urease test were suitable samples for clarithromycin resistance detection in patients infected with H. pylori, which became especially useful in cases where the number or size of the biopsies is limited. In addition, this is the first report of a molecular analysis for clarithromycin resistance performed directly from gastric biopsies in our region.

摘要

幽门螺杆菌是一种胃部病原体,被广泛认为是胃部疾病的致病因子。其根除情况存在差异,主要是由于对克拉霉素的耐药性增加。我们的目的是:评估(i)用于快速尿素酶试验的活检标本是否是通过聚合酶链反应-限制性片段长度多态性(PCR-RFLP)检测对克拉霉素耐药性的有用样本,以及(ii)23S rRNA基因中A2142G和A2143G点突变的分布与我们地区毒力因子的关系。从成年消化不良患者(n = 141)中收集胃标本,并通过快速尿素酶试验、组织病理学分析和hsp60基因的PCR检测幽门螺杆菌。在62份提交进行分子分析和快速尿素酶试验的幽门螺杆菌(+)配对活检标本中,通过PCR-RFLP检测克拉霉素耐药性。使用针对cagA、vacA和babA2基因的特异性引物,通过多重PCR分析幽门螺杆菌毒力因子。62株菌株中有13株(20.9%)对克拉霉素耐药:6/13(46.2%)携带A2143G突变,而7/13(53.8%)携带A2142G点突变。vacA m1s1是耐药菌株中最常见的基因型。总之,用于快速尿素酶试验的活检标本是检测幽门螺杆菌感染患者克拉霉素耐药性的合适样本,在活检数量或大小有限的情况下尤其有用。此外,这是我们地区首次直接从胃活检标本进行克拉霉素耐药性分子分析的报告。

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