Journal List > Korean J Clin Microbiol > v.11(2) > 1038156

Lee, Chae, Kang, Lee, Sung, Kim, Lee, Lee, and Shim: Multicenter Study for the Frequency of 23S rRNA Point Mutations Associated with Clarithromycin Resistance in Helicobacter pylori in Korea

Abstract

Background

Clarithromycin resistance in Helicobacter pylori is a major cause of eradication therapy failure. The objective of this study was to determine the frequency and type of mutations in the 23S rRNA gene in Korea, which are associated with clarithromycin resistance.

Methods

From January 2008 to March 2008, 353 gastric biopsy specimens were collected from five university hospitals in Seoul and Kyunggido. H. pylori infection was defined as showing a positive result in at least one of the following three tests: a microaerophilic culture, a CLO test, and a Giemsa/silver stain. The frequencies of A2143G, A2142G, and the wild type of 23S rRNA and the presence of H. pylori were determined by Seeplex ClaR-H. pylori PCR (Seegene Inc., Seoul, Korea). Twenty-nine culture isolates were tested for susceptibility to clarithromycin by E-test (AB Biodisk, Solna, Sweden) or the CLSI (Clinical and Laboratory Standards Institute) disk diffusion test.

Results

From 176 H. pylori PCR-positive specimens, 23S rRNA gene mutations were detected in 38 isolates (21.6%), including 27 isolates of A2143G and 11 isolates of A2142G. Total mutation rates varied from 15.8% to 31.3% with the frequency of A2143G mutation alone varying from 8.5% to 25.0% among the five hospitals studied. There were 10 clarithromycin-resistant isolates found by susceptibility test and they were all positive for A2143G mutation. But, 3 of the 19 susceptible isolates were also positive for either A2143G or A2142G mutation.

Conclusion

In Korea, the overall frequency of clarithromycin-resistant H. pylori was 21.6%; however, the type and frequency of the 23S rRNA mutations varied from hospital to hospital.

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Fig. 1.
Detection of the point mutations at 2143 (A to G) and 2142 (A to G) in 23S rRNA gene by polymerase chain reaction products. Lane M, DNA ladder; lane 1–3 and 5–7, wild type; lane 4, mutant type of A2142G.
kjcm-11-84f1.tif
Table 1.
Endoscopic diagnosis of patients at 5 university hospitals in Korea
Hospital Gastritis Gastric ulcer Gastric polyp/adenoma Gastric cancer Total
A 18 8 0 10 36
B 76 13 0 4 93
C 31 3 0 0 34
D 77 20 0 3 100
E 69 13 6 2 90
Total 271 57 6 19 353
Table 2.
Comparison of Seeplex ClaR-H. pylori PCR and other methods
Seeplex ClaR-H. pylori PCR CLO or Giemsa/silver stain or culture
Positive Negative
Positive 163 13
Negative 31 146
Total 194 159

Abbreviations: ClaR-H. pylori, clarithromycin-resistant Helicobacter pylori

The concordance rate of Seeplex ClaR-H. pylori PCR and other methods was 87.5% (309/353).

Table 3.
Allele frequencies of A2143G and A2142G at five university hospitals in Korea
Hospital Wild type Mutations (%) of 23S rRNA gene
A2142G A2143G Total
A 30 4 (9.5)0 8 (19.0) 12 (28.5)
B 32 0 (0.0)0 6 (15.8) 6 (15.8)
C 48 6 (10.1) 5 (8.5)0 11 (18.6)
D 11 1 (6.3)0 4 (25.0) 5 (31.3)
E 17 0 (0.0)0 4 (19.0) 4 (19.0)
Total 138 11 (6.3)0 27 (15.3) 38 (21.6)
Table 4.
Comparison of genotype and phenotype to detect clarithromycin resistance
23S rRNA genotype Phenotype of clarithromycin susceptibility
Susceptible (n=19) Resistant (n=10)
Wild type 16 0
A2143G 1 10
A2142G 2 0
Total 19 10
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