Detection of four clarithromycin resistance point mutations in Helicobacter pylori: comparison of real-time PCR and PCR-RFLP methods

2012 ◽  
Vol 22 (5) ◽  
pp. 1007-1013 ◽  
Author(s):  
Mohammad Kargar ◽  
Abbas Doosti ◽  
Sadegh Ghorbani-Dalini
Keyword(s):  
Helicobacter Pylori ◽  
Real Time ◽  
Real Time Pcr ◽  
Point Mutations ◽  
Clarithromycin Resistance ◽  
Pcr Rflp

Change of point mutations in Helicobacter pylori rRNA associated with clarithromycin resistance in Italy

2014 ◽  
Vol 63 (3) ◽  
pp. 453-457 ◽  
Author(s):  
Vincenzo De Francesco ◽  
Angelo Zullo ◽  
Floriana Giorgio ◽  
Ilaria Saracino ◽  
Cristina Zaccaro ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Real Time ◽  
Real Time Pcr ◽  
Single Point ◽  
Point Mutations ◽  
Clarithromycin Resistance ◽  
Genotypic Analysis ◽  
Previously Treated ◽  
Resistant Strains ◽  
H Pylori

Primary clarithromycin resistance is the main factor affecting the efficacy of Helicobacter pylori therapy. This study aimed: (i) to assess the concordance between phenotypic (culture) and genotypic (real-time PCR) tests in resistant strains; (ii) to search, in the case of disagreement between the methods, for point mutations other than those reported as the most frequent in Europe; and (iii) to compare the MICs associated with the single point mutations. In order to perform real-time PCR, we retrieved biopsies from patients in whom H. pylori infection was successful diagnosed by bacterial culture and clarithromycin resistance was assessed using the Etest. Only patients who had never been previously treated, and with H. pylori strains that were either resistant exclusively to clarithromycin or without any resistance, were included. Biopsies from 82 infected patients were analysed, including 42 strains that were clarithromycin resistant and 40 that were clarithromycin susceptible on culture. On genotypic analysis, at least one of the three most frequently reported point mutations (A2142C, A2142G and A2143G) was detected in only 23 cases (54.8 %), with a concordance between the two methods of 0.67. Novel point mutations (A2115G, G2141A and A2144T) were detected in a further 14 out of 19 discordant cases, increasing the resistance detection rate of PCR to 88 % (P<0.001; odds ratio 6.1, 95 % confidence interval 2−18.6) and the concordance to 0.81. No significant differences in MIC values among different point mutations were observed. This study suggests that: (i) the prevalence of the usually reported point mutations may be decreasing, with a concomitant emergence of new mutations; (ii) PCR-based methods should search for at least six point mutations to achieve good accuracy in detecting clarithromycin resistance; and (iii) none of the tested point mutations is associated with significantly higher MIC values than the others.

HELICOBACTER PYLORI CLARITHROMYCIN RESISTANCE ASSESSMENT BY CULTURE AND TAQMAN REAL-TIME PCR: A COMPARATIVE STUDY

2009 ◽  
Vol 41 ◽  
pp. S85 ◽  
Author(s):  
V. De Francesco ◽  
A. Zullo ◽  
F. Perna ◽  
F. Giorgio ◽  
C. Hassan ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Comparative Study ◽  
Real Time ◽  
Real Time Pcr ◽  
Clarithromycin Resistance ◽  
Resistance Assessment

scholarly journals PCR-RFLP detection of point mutations A2143G and A2142G in 23S rRNA gene conferring resistance to clarithromycin in Helicobacter pylori strains.

2014 ◽  
Vol 61 (2) ◽  
Author(s):  
Karolina Klesiewicz ◽  
Paweł Nowak ◽  
Elżbieta Karczewska ◽  
Iwona Skiba ◽  
Izabela Wojtas-Bonior ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Point Mutations ◽  
Rflp Analysis ◽  
23S Rrna ◽  
Rrna Gene ◽  
Efficient Management ◽  
Clarithromycin Resistance ◽  
Pcr Rflp ◽  
23S Rrna Gene ◽  
H Pylori

The occurrence of clarithromycin resistance among Helicobacter pylori strains is a major cause of the treatment failure. Resistance to this drug is conferred by point mutations in 23S rRNA gene and the most prevalent mutations are A2143G and A2142G. The aim of the study was to evaluate the occurrence of A2143G and A2142G mutations in a group of H. pylori strains resistant to clarithromycin. The study included 21 clarithromycin-resistant H. pylori strains collected between 2006 and 2009 in southern Poland. Resistance to clarithromycin was quantitatively tested with the E-test to determine the minimal inhibitory concentration (MIC value). The point mutations of H. pylori isolates were detected by PCR followed by RFLP analysis. The MIC values for clarithromycin for the analyzed strains ranged from 1.5 mg/L to 64 mg/L. Nine H. pylori strains exhibited A2143G mutation and A2142G mutation was found in 9 isolates as well. The results of RFLP analysis of 3 clarithromycin-resistant strains were negative for both mutations. The average MIC values for A2143G and A2142G mutants were 6 and 30 mg/L, respectively. Frequencies of A2143G and A2142G mutations were the same in all isolates tested. Strains with A2143G mutation exhibited lower MIC values than A2142G mutants. Application of PCR-RFLP method for detection of clarithromycin resistance allows for better and more efficient management of H. pylori infections.

Routine use of real-time PCR for detection of Helicobacter pylori and of clarithromycin resistance mutations

2007 ◽  
Vol 31 (10) ◽  
pp. 792-795 ◽  
Author(s):  
Jacques Tankovic ◽  
Marie-Thérèse Chaumette-Planckaert ◽  
Lionel Deforges ◽  
Nathalie Launay ◽  
Jeanne-Marie Le Glaunec ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Real Time ◽  
Real Time Pcr ◽  
Resistance Mutations ◽  
Clarithromycin Resistance

PCR-RFLP DETECTION OF POINT MUTATIONS CONFERRING RESISTANCE TO CLARITHROMYCIN OF HELICOBACTER PYLORI IN QUANG NGAI

2015 ◽  
pp. 54-61
Author(s):  
Ngoc Doanh Pham ◽  
Van Huy Tran ◽  
Thi Minh Thi Ha
Keyword(s):  
Helicobacter Pylori ◽  
General Hospital ◽  
Chronic Gastritis ◽  
Point Mutations ◽  
23S Rrna ◽  
Rrna Gene ◽  
Clarithromycin Resistance ◽  
Urease Test ◽  
Pcr Rflp ◽  
H Pylori

Background: Clarithromycin resistance of H-pylori is the main cause leading to treatment failure. Aim: The purpose of this study was to determine the rate of clarithromycin resistance mutation on gene 23S ribosomal popular robonucleotide acid (rRNA) of H-pylori in patients with chronic gastritis in Quang Ngai General Hospital PCR-RFLP. Method: This is a cross-sectional study in 64 patients infected with H-pylori was determined by 3 methods and chronic gastritis proven by histology. Sample collection conducted in Quang Ngai general hospital and molecular biology tests were conducted in the medical genetics department Hue of Medical and Pharmaceutical University. Urease test, histopathological examination and perform HE staining PCR 23S rRNA gene fragment of H-pylori to determine H-pylori infection. Analysis of genetic mutations in the 23S rRNA point is performed by PCR-RFLP technique. Results: Of the 64 biopsies qualify included in the study, 41 samples with clarithromycin resistance point mutations (64%), of which 40 (62.5%) had mutations A2143A, one sample with A2142A (2%). No samples had mutations A2142C and no more than one mutation.Conclusion: This is the first time we report mutations related to clarithromycin of H-pylori in Quang Ngai province. Mutations rate is high (64%), among the common mutations, the most common mutantation is A2143G. Key words: Helicobacter pylori, clarithromycin resistance, PCR-RFLP, point mutations

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