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使用基于重复序列的聚合酶链反应的DiversiLab微生物分型系统在日本对幽门螺杆菌进行特征分析的临床应用

Clinical Application of the DiversiLab Microbial Typing System Using Repetitive Sequence-Based PCR for Characterization of Helicobacter pylori in Japan.

作者信息

Morimoto Norihito, Takeuchi Hiroaki, Nishida Yoshie, Morisawa Mie, Yoshikawa Tomoe, Morita Tamae, Morimoto Miyuki, Sugimoto Chizuko, Matsumura Yoshihisa, Sugiura Tetsuro

机构信息

Department of Clinical Laboratory, Kochi Medical School Hospital, Kochi University, Kohasu, Oko-cho, Nankoku, Kochi, Japan.

Clinical Laboratory Medicine, Kochi University School of Medicine, Kochi, Japan.

出版信息

J Clin Lab Anal. 2015 May;29(3):250-3. doi: 10.1002/jcla.21758. Epub 2014 May 5.

Abstract

We evaluated the DiversiLab (DL) system with universal primers, a semiautomated repetitive extragenic palindromic sequence-based polymerase chain reaction (PCR) (rep-PCR) system, for the characterization of Helicobacter pylori in Japan. All 135 isolates from Japanese patients with gastric cancer (GC, n = 55) or non-GC (n = 80) were used and subjected to the drug susceptibility examinations (amoxicillin, AMPC; metronidazole, MNZ; and clarithromycin, CAM) by E-test. There were 28 MNZ-resistant (20.7%), 35 CAM-resistant (25.9%), and 16 MNZ/CAM-resistant (11.9%) isolates. DL rep-PCR fingerprinting analysis at the level of 95% similarity revealed five major groups (A-E) and the other including 45 isolates. The occupation rates of GC-derived isolates in groups B (54.2%) and E (58.8%) were higher than in the other groups: A (26.7%), C (28.6%), D (30.0%), and the other (40.0%). Relative higher occupation rates of drug resistants, such as MNZ-, CAM- and double MNZ/CAM-resistant isolates, were observed in groups B (45.8%), C (42.6%), and D (40%). Five of eight GC-derived isolates with MNZ/CAM resistance were significantly assigned to group B (P = 0.0312, χ(2) -test). These results suggest that the isolates classified in group B have a potential to contribute to the development of severe gastric disorders. The DL system, rapid and high sensitive technology, would be widely available in clinical laboratory for pathological and epidemiological analyses even in H. pylori.

摘要

我们使用通用引物对DiversiLab(DL)系统进行了评估,该系统是一种基于半自动化重复基因外回文序列的聚合酶链反应(PCR)(rep-PCR)系统,用于对日本幽门螺杆菌进行特征分析。我们使用了来自日本胃癌(GC,n = 55)或非GC(n = 80)患者的所有135株分离株,并通过E-test进行药敏试验(阿莫西林,AMPC;甲硝唑,MNZ;克拉霉素,CAM)。有28株对MNZ耐药(20.7%),35株对CAM耐药(25.9%),16株对MNZ/CAM耐药(11.9%)。DL rep-PCR指纹图谱分析在95%相似性水平上揭示了五个主要组(A-E)以及包括45株分离株的其他组。B组(54.2%)和E组(58.8%)中GC来源分离株的占比高于其他组:A组(26.7%)、C组(28.6%)、D组(30.0%)和其他组(40.0%)。在B组(45.8%)、C组(42.6%)和D组(40%)中观察到耐药菌(如对MNZ、CAM和双重MNZ/CAM耐药的分离株)的相对较高占比。八株对MNZ/CAM耐药的GC来源分离株中有五株显著归为B组(P = 0.0312,χ²检验)。这些结果表明,归为B组的分离株有可能导致严重胃部疾病的发生。DL系统是一种快速且高灵敏度的技术,即使在幽门螺杆菌检测中,也将在临床实验室中广泛用于病理和流行病学分析。

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