1887

Journal of Medical Microbiology logo

Volume 58, Issue 11

Enzymic colorimetry-based DNA chip: a rapid and accurate assay for detecting mutations for clarithromycin resistance in the 23S rRNA gene of Free

Abstract

Macrolide drugs, such as clarithromycin (CAM), are a key component of many combination therapies used to eradicate . However, resistance to CAM is increasing in and is becoming a serious problem in eradication therapy. CAM resistance in is mostly due to point mutations (A2142G/C, A2143G) in the peptidyltransferase-encoding region of the 23S rRNA gene. In this study an enzymic colorimetry-based DNA chip was developed to analyse single-nucleotide polymorphisms of the 23S rRNA gene to determine the prevalence of mutations in CAM-related resistance in -positive patients. The results of the colorimetric DNA chip were confirmed by direct DNA sequencing. In 63 samples, the incidence of the A2143G mutation was 17.46 % (11/63). The results of the colorimetric DNA chip were concordant with DNA sequencing in 96.83 % of results (61/63). The colorimetric DNA chip could detect wild-type and mutant signals at every site, even at a DNA concentration of 1.53×10 copies μl. Thus, the colorimetric DNA chip is a reliable assay for rapid and accurate detection of mutations in the 23S rRNA gene of that lead to CAM-related resistance, directly from gastric tissues.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): CAM, clarithromycin , NC, negative control , PC, positive control and SNP, single-nucleotide polymorphism
SGM
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.010785-0
2009-11-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/jmm/58/11/1443.html?itemId=/content/journal/jmm/10.1099/jmm.0.010785-0&mimeType=html&fmt=ahah

References

  1. Bener A., Uduman S. A., Ameen A., Alwash R., Pasha M. A., Usmani M. A., Al-Naili S. R., Amiri K. M. 2002; Prevalence of Helicobacter pylori infection among low socio-economic workers. J Commun Dis 34:179–184
     [Google Scholar]
  2. Buzás G. M., Lotz G., Kiss A. 2007; The epidemiology of clarithromycin resistance of Helicobacter pylori infection in Hungary. Orv Hetil 148:1461–1467 [CrossRef]
     [Google Scholar]
  3. Cheng H., Hu F. L. 2005; The epidemiology of Helicobacter pylori resistance to antibiotics in Beijing. Zhonghua Yi Xue Za Zhi 85:2754–2757 (in Chinese
    [Google Scholar]
  4. Chizhikov V., Wagner M., Ivshina A., Hoshino Y., Kapikian A. Z., Chumakov K. 2002; Detection and genotyping of human group A rotaviruses by oligonucleotide microarray hybridization. J Clin Microbiol 40:2398–2407 [CrossRef]
     [Google Scholar]
  5. Drumm B., Koletzko S., Oderda G. 2000; Helicobacter pylori infection in children: a consensus statement. European Paediatric Task Force on Helicobacter pylori . J Pediatr Gastroenterol Nutr 30:207–213 [CrossRef]
     [Google Scholar]
  6. Dunn B. E., Cohen H., Blaser M. J. 1997; Helicobacter pylori . Clin Microbiol Rev 10:720–741
     [Google Scholar]
  7. Fresnadillo Martínez M. J., Rodríguez Rincón M., Blázquez de Castro A. M., García Sánchez E., García Sánchez J. E., Trujillano Martín I., Cordero Sánchez M., Alvarez Alvarez P., Paz Bouza J., García-Rodríguez J. A. 1997; Comparative evaluation of selective and nonselective media for primary isolation of Helicobacter pylori from gastric biopsies. Helicobacter 2:36–39 [CrossRef]
     [Google Scholar]
  8. Hao Q., Li Y., Zhang Z. J., Liu Y., Gao H. 2004; New mutation points in 23S rRNA gene associated with Helicobacter pylori resistance to clarithromycin in northeast China. World J Gastroenterol 10:1075–1077
     [Google Scholar]
  9. Heep M., Scheibl K., Degrell A., Lehn N. 1999; Transport and storage of fresh and frozen gastric biopsy specimens for optimal recovery of Helicobacter pylori . J Clin Microbiol 37:3764–3766
     [Google Scholar]
  10. Kato S., Fujimura S., Udagawa H., Shimizu T., Maisawa S., Ozawa K., Iinuma K. 2002; Antibiotic resistance of Helicobacter pylori strains in Japanese children. J Clin Microbiol 40:649–653 [CrossRef]
     [Google Scholar]
  11. Kim K. S., Kang J. O., Eun C. S., Han D. S., Choi T. Y. 2002; Mutations in the 23S rRNA gene of Helicobacter pylori associated with clarithromycin resistance. J Korean Med Sci 17:599–603 [CrossRef]
     [Google Scholar]
  12. Kim J. M., Kim J. S., Jung H. C., Kim N., Kim Y. J., Song I. S. 2004; Distribution of antibiotic MICs for Helicobacter pylori strains over a 16-year period in patients from Seoul, South Korea. Antimicrob Agents Chemother 48:4843–4847 [CrossRef]
     [Google Scholar]
  13. Kobayashi I., Saika T., Muraoka H., Murakami K., Fujioka T. 2006; Helicobacter pylori isolated from patients who later failed H. pylori eradication triple therapy readily develop resistance to clarithromycin. J Med Microbiol 55:737–740 [CrossRef]
     [Google Scholar]
  14. Liu Z., Shen J., Zhang L., Shen L., Li Q., Zhang B., Zhou J., Gu L., Feng G. other authors 2008; Prevalence of A2143G mutation of H. pylori -23S rRNA in Chinese subjects with and without clarithromycin use history. BMC Microbiol 8:81 [CrossRef]
     [Google Scholar]
  15. Maeda S., Yoshida H., Matsunaga H., Ogura K., Kawamata O., Shiratori Y., Omata M. 2000; Detection of clarithromycin-resistant Helicobacter pylori strains by a preferential homoduplex formation assay. J Clin Microbiol 38:210–214
     [Google Scholar]
  16. Mao H., Wang H., Zhang D., Mao H., Zhao J., Shi J., Cui Z. 2006; Study of hepatitis B virus gene mutations with enzymatic colorimetry-based DNA microarray. Clin Biochem 39:67–73 [CrossRef]
     [Google Scholar]
  17. Mégraud F. 2004; H. pylori antibiotic resistance: prevalence, importance, and advances in testing. Gut 53:1374–1384 [CrossRef]
     [Google Scholar]
  18. Pan Z. J., Su W. W., Tytgat G. N., Dankert J., Van der Ende A. 2002; Assessment of clarithromycin-resistant Helicobacter pylori among patients in Shanghai and Guangzhou, China, by primer-mismatch PCR. J Clin Microbiol 40:259–261 [CrossRef]
     [Google Scholar]
  19. Rüssmann H., Adler K., Haas R., Gebert B., Koletzko S., Heesemann J. 2001; Rapid and accurate determination of genotypic clarithromycin resistance in cultured Helicobacter pylori by fluorescent in situ hybridization. J Clin Microbiol 39:4142–4144 [CrossRef]
     [Google Scholar]
  20. Sezgin O., Aslan G., Altintaş E., Tezcan S., Serin M. S., Emekdaş G. 2008; Detection of point mutations on 23S rRNA of Helicobacter pylori and resistance to clarithromycin with PCR-RFLP in gastric biopsy specimens in Mersin, Turkey. Turk J Gastroenterol 19:163–167
     [Google Scholar]
  21. Strnad M., Presecki V., Babus V., Turek S., Dominis M., Kalenić S., Hebrang A., Katicić M. 2002; Epidemiology of Helicobacter pylori infection. Lijec Vjesn 124 (Suppl. 1):5–9
     [Google Scholar]
  22. Sung J. J., Chung S. C., Ling T. K., Yung M. Y., Leung V. K., Ng E. K., Li M. K., Cheng A., Li A. K. 1995; Antibacterial treatment of gastric ulcers associated with Helicobacter pylori . N Engl J Med 332:139–142 [CrossRef]
     [Google Scholar]
  23. Taylor D. E., Ge Z., Purych D., Lo T., Hiratsuka K. 1997; Cloning and sequence analysis of two copies of a 23S rRNA gene from Helicobacter pylori and association of clarithromycin resistance with 23S rRNA mutations. Antimicrob Agents Chemother 41:2621–2628
     [Google Scholar]
  24. Treiber G., Kist M., Klotz U., Peitz U., Malfertheiner P. 2005; Therapie der Helicobacter -pylori-infektion: klinische, mikrobiologische und pharmakologische aspekte. Dtsch Arztebl 26:1883–1888 (in German
    [Google Scholar]
  25. Van Doorn L. J., Glupczynski Y., Kusters J. G., Me'graud F., Midolo P., Maggi-Solca N., Queiroz D. M. M., Nouhan N., Stet E., Quint W. G. V. 2001; Accurate prediction of macrolide resistance in Helicobacter pylori by a PCR line probe assay for detection of mutations in the 23S rRNA gene: multicenter validation study. Antimicrob Agents Chemother 45:1500–1504 [CrossRef]
     [Google Scholar]
  26. Xing J. Z., Clarke C., Zhu L., Gabos S. 2005; Development of a microelectronic chip array for high-throughput genotyping of Helicobacter species and screening for antimicrobial resistance. J Biomol Screen 10:235–245 [CrossRef]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.010785-0
Loading
Enzymic colorimetry-based DNA chip: a rapid and accurate assay for detecting mutations for clarithromycin resistance in the 23S rRNA gene of Helicobacter pylori
J. Med. Microbiol. 58, 1443 (2009); https://doi.org/10.1099/jmm.0.010785-0
/content/journal/jmm/10.1099/jmm.0.010785-0
/content/journal/jmm/10.1099/jmm.0.010785-0
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error