Detection ofHelicobacter pylori by polymerase chain reaction assay using gastric biopsy specimens taken for CLOtest

1996 ◽  
Vol 31 (3) ◽  
pp. 329-332 ◽  
Author(s):  
Tin-Tsan Lin ◽  
Chau-Ting Yeh ◽  
Ellen Yang ◽  
Pang-Chi Chen
Keyword(s):  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Assay ◽  
Gastric Biopsy ◽  
Chain Reaction ◽  
Biopsy Specimens ◽  
Polymerase Chain

scholarly journals Direct detection of Helicobacter pylori resistance to macrolides by a polymerase chain reaction/DNA enzyme immunoassay in gastric biopsy specimens

Gut ◽  
1999 ◽  
Vol 44 (4) ◽  
pp. 463-467 ◽  
Author(s):  
A Marais ◽  
L Monteiro ◽  
A Occhialini ◽  
M Pina ◽  
H Lamouliatte ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Gastric Biopsy ◽  
23S Rrna ◽  
Wild Type ◽  
Chain Reaction ◽  
Biopsy Specimens ◽  
Resistant Strains ◽  
Polymerase Chain ◽  
H Pylori

BACKGROUNDThe increasing use of macrolides especially in the treatment ofHelicobacter pylori infection has led to an increase in resistant strains. The resistance of H pylori to macrolides, especially clarithromycin, is one of the major causes of eradication failure. In H pylori, clarithromycin resistance is due to point mutations localised in domain V of 23S rRNA.AIMTo develop a molecular technique based on amplification of a relevant fragment of the 23S rRNA and colorimetric hybridisation in liquid phase to detect directly in biopsy specimens the type of mutation associated with resistance of H pylori to clarithromycin.METHODSGastric biopsy samples from 61 patients were submitted to this test. The results were compared with standard methods (determination of minimal inhibition concentration, polymerase chain reaction/restriction fragment length polymorphism, and/or DNA sequencing) in order to evaluate the test and to define the cut off values, specificity, and sensitivity.RESULTSThe 14 biopsy samples in which H pylori was not detected did not give a positive result in any assay, and the 14 samples harbouring strains susceptible to clarithromycin gave a positive result with the wild type probe as expected. The 33 biopsy specimens containing resistant strains always gave a positive signal with one of the probes detecting resistant organisms, but in eight cases they also reacted with the wild type probe, indicating that a mixture of resistant and susceptible organisms was present.CONCLUSIONThe importance of this new assay is that it allows the detection of multiple genotypes corresponding to either heterogeneous genotypes or mixed infections. Moreover, it allows in a single step not only the detection of H pylori but also the determination of its susceptibility to clarithromycin directly in biopsy specimens without the need for culture.

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