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Journal of Clinical Microbiology, July 2003, p. 3387-3391, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.3387-3391.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Detection of Helicobacter pylori in Gastric Mucosa of Patients with Gastroduodenal Diseases by PCR-Restriction Analysis Using the RNA Polymerase Gene (rpoB)

Chang-Young Lim,1,{dagger} Keun-Hwa Lee,2 Myung-Je Cho,3 Myung-Woong Chang,4 Seok-Yong Kim,5 Na-Hye Myong,6 Woo-Kon Lee,3 Kwang-Ho Rhee,3 and Yoon-Hoh Kook2*

Department of Internal Medicine,1 Department of Pathology, Dankook University College of Medicine, Cheonan, 330-715,6 Department of Microbiology, Institute of Endemic Diseases, SNUMRC, and Cancer Research Center, Seoul National University College of Medicine, and Clinical Research Institute, Seoul National University Hospital, Seoul 110-799,2 Department of Microbiology, Gyeong-Sang National University College of Medicine,3 Department of Microbiology, School of Medicine, Kosin University, Busan 602-702,4 Department of Microbiology, College of Medicine, Chungbuk National University, Cheongju 361-711, Korea5

Received 11 December 2002/ Returned for modification 26 March 2003/ Accepted 21 April 2003

A novel PCR restriction analysis method using the RNA polymerase ß-subunit- coding gene (rpoB) was employed to both detect and identify Helicobacter pylori in biopsy specimens and culture isolates. The rpoB DNAs (458 bp) were specifically amplified by PCR with the Helicobacter-specific primers (HF and HR). Based on the determined rpoB sequences of the culture isolates, an H. pylori-specific restriction site, Tru9I, was found. H. pylori can be identified by observing two discernible DNA fragments (288 and 138 bp) after Tru9I digestion and agarose gel electrophoresis. The rpoB PCR and subsequent restriction analysis (PRA) enabled the specific detection and identification of H. pylori in biopsy specimens from patients with gastroduodenal diseases. The rpoB PRA conferred a compatible or a slightly higher positive rate (53.7%) than did the Campylobacter-like organism (CLO) test (50.4%) and glmM PCR (48.8%), suggesting that it is useful for diagnosing an H. pylori infection without culture in the clinical laboratory.

* Corresponding author. Mailing address: Department of Microbiology, Seoul National University College of Medicine, 28 Yongon-dong, Chongno-gu, Seoul 110-799, Korea. Phone: (82) 2-740-8306. Fax: (82) 2-743-0881. E-mail: yhkook{at}plaza.snu.ac.kr.

{dagger} Present address: Department of Internal Medicine, Hansol Hospital, Songpagu, Seoul 138-844, Korea.

Journal of Clinical Microbiology, July 2003, p. 3387-3391, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.3387-3391.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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