Annals of Laboratory Medicine

Korean Society for Laboratory Medicine (대한진단검사의학회)
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A Comparison Analysis on the Diagnosis of Helicobacter pylori Infection and the Detection of Clarithromycin Resistance according to Biopsy Sites

  • Cho, Ah-Ra (Department of Laboratory Medicine, Chung-Ang University College of Medicine) ;
  • Lee, Mi-Kyung (Department of Laboratory Medicine, Chung-Ang University College of Medicine)
  • Received : 2010.02.12
  • Accepted : 2010.07.26
  • Published : 2010.08.31
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Abstract

Background : This study was performed to determine the biopsy sites that are suitable for the diagnosis of Helicobacter pylori infection and to assess the sensitivity of culture, histology, and dual-priming oligonucleotide (DPO)-based multiplex PCR. Moreover, we evaluated the usefulness of PCR for the detection of 23S rRNA mutations, which are responsible for the clarithromycin resistance of H. pylori. Methods : From 90 patients, we obtained biopsy specimens for culture, histology, and Seeplex.. ClaR-H. pylori PCR (Seegene Inc., Korea). Phenotypic susceptibility to clarithromycin was evaluated using the E-test (AB Biodisk, Sweden). Results : H. pylori was detected in 48 of 90 patients. The positive rates of infection in the antrum and body were higher than those in the biopsies obtained from the duodenal bulb. The positive rates in histology, PCR, and culture were 46.7%, 42.2%, and 34.4%, respectively. Using histology or PCR, we identified H. pylori in 46 of the 48 patients. 23S rRNA mutations were detected in 8 patients. The clarithromycin E-test showed that all the 10 wild-type patients were susceptible. However, the results of the PCR and E-test of 3 of the 8 mutation-positive patients were discrepant. Conclusions : We observed that a combination of histology and PCR affords a high detection rate of H.pylori infection and that DPO-based PCR can be practically used for the diagnosis of H. pylori infection and the determination of clarithromycin resistance. These techniques were useful for biopsy sampling simultaneously from the antrum and body for the detection of clarithromycin resistance of multiple strain infection or heteroresistance.

배경 : 본 연구에서는 위 생검부위에 따른 H. pylori 검출률을 관찰하였으며 배양, 조직검사 및 dual priming oligonucleotide (DPO)-based multiplex PCR을 실시하여 방법간의 민감도를 비교하였다. 또한 clarithromycin 내성과 연관된 23S rRNA 돌연변이를 검출하기 위한 multiplex PCR의 유용성을 평가하였다. 방법 : 90명의 환자에게 위 전정부, 체부 및 십이지장 구부에서 생검한 조직을 배양, 조직검사 및 Seeplex ClaR-H. pylori PCR (Seegene Inc., Korea)을 시행하였다. Clarithromycin 내성 표현형 검사는 E-test (AB Biodisk, Sweden)로 시행하였다. 결과 : 90명 환자 중 48명(53%)이 H. pylori 양성이었다. 위전정부와 체부에 비해 십이지장 구부는 낮은 검출률을 보였다. 조직검사 46.7% (42/90), PCR 42.2% (38/90), 배양 34.4% (31/90)로 조직검사의 양성률이 제일 높았으며 조직검사와 PCR을 동시에 했을 95.8% (46/48)로 거의 모든 환자를 검출할 수 있었다. DPO-based multiplex PCR에서는 8명의 돌연변이 환자가 검출되었다. Clarithromycin 내성 E-test에서는 유전형에서 wild type로 나온 10명의 23균주는 모두 감수성을 보였으나 돌연변이 3명은 표현형과 유전형이 불일치하였다. 결론 : H. pylori는 조직검사와 PCR을 같이 시행하였을 경우 높은 검출률을 보이며 DPO-based multiplex PCR를 시행할 경우 PCR과 내성 검사를 동시에 시행할 수 있어 임상에 유용한 정보를 제공할 수 있다. 또한 위전정부와 체부를 동시에 검사함으로써 혼합감염 및 hetero-resistance 균주로 인한 clarithromycin 내성을 검출할 수 있겠다.

Keywords

Acknowledgement

Supported by : Chung-Ang University

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