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Validation of a PNA-FISH method for the detection of Helicobacter pylori clarithromycin resistance in gastric biopsies

Validation of a PNA-FISH method for the detection of Helicobacter pylori clarithromycin resistance in gastric biopsies
Validation of a PNA-FISH method for the detection of Helicobacter pylori clarithromycin resistance in gastric biopsies
In here, we evaluated a previously established PNA-FISH method as a new diagnostic test for H. pylori clarithromycin resistance detection in paraffin embedded gastric biopsies. Both a retrospective and a prospective cohort study were conducted to evaluate the specificity and sensitivity of a PNA-FISH method to determine H. pylori clarithromycin resistance. In the retrospective study (n=30 patients), full agreement between PNA-FISH and PCR-sequencing was observed. When compared to the reference method (culture followed by Etest®), specificity and sensitivity of PNA-FISH were 90.9% (95% confidence interval [CI], 57.1-99.5%) and 84.2% (95% CI, 59.5-95.8%), respectively. In the prospective cohort (n=93 patients), 21 cases were positive by culture. For the patients harboring clarithromycin-resistant H. pylori, the method showed a sensitivity of 80.0% (95% CI, 29.9-98.9%) and specificity of 93.8% (95% CI, 67.7-99.7%). These values are likely to be underestimated, as in some discrepant results patients were infected by more than one strain. PNA-FISH appears to be a simple, quick and an accurate method for detecting
H. pylori clarithromycin resistance in paraffin embedded biopsies. It is also the only one of the methods assessed here that allows direct and specific visualization of this microorganism within the biopsies, a characteristic that allowed observing that different H. pylori strains can subsist very closely in the stomach.
0095-1137
Cerqueira, Laura
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Fernandes, Ricardo M.
8c788439-4904-486f-af7b-189855fb00da
Ferreira, Rui M.
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Oleastro, Mónica
e4b443f4-aace-4ef2-be40-69f79ec81d6b
Carneiro, Fátima
09da70a4-8ca4-4078-b177-f3ccd870b447
Brandao, Catarina
b830bbb8-432e-47ba-90ff-493b9ad6fe53
Pimentel-Nunes, Pedro
0075ecf3-822f-4dc8-8e61-5b5790da2f21
Dinis-Ribeiro, Mário
7d966cbc-870e-43cd-a3c4-6685599d27bd
Figueiredo, Céu
54d5e121-a8e1-48f1-93d0-453b7fa9a97f
Keevil, Charles W.
cb7de0a7-ce33-4cfa-af52-07f99e5650eb
Vieira, Maria J.
d972e877-d85b-488c-8b0f-358f79d2fa29
Azevedo, Nuno F.
24c4eb52-0c98-443b-881f-7a1449c9ac26
Cerqueira, Laura
61343b41-4742-4057-94e9-2ef4ea145613
Fernandes, Ricardo M.
8c788439-4904-486f-af7b-189855fb00da
Ferreira, Rui M.
8ec38033-099b-43e3-80b9-2c736b6e3394
Oleastro, Mónica
e4b443f4-aace-4ef2-be40-69f79ec81d6b
Carneiro, Fátima
09da70a4-8ca4-4078-b177-f3ccd870b447
Brandao, Catarina
b830bbb8-432e-47ba-90ff-493b9ad6fe53
Pimentel-Nunes, Pedro
0075ecf3-822f-4dc8-8e61-5b5790da2f21
Dinis-Ribeiro, Mário
7d966cbc-870e-43cd-a3c4-6685599d27bd
Figueiredo, Céu
54d5e121-a8e1-48f1-93d0-453b7fa9a97f
Keevil, Charles W.
cb7de0a7-ce33-4cfa-af52-07f99e5650eb
Vieira, Maria J.
d972e877-d85b-488c-8b0f-358f79d2fa29
Azevedo, Nuno F.
24c4eb52-0c98-443b-881f-7a1449c9ac26

Cerqueira, Laura, Fernandes, Ricardo M., Ferreira, Rui M., Oleastro, Mónica, Carneiro, Fátima, Brandao, Catarina, Pimentel-Nunes, Pedro, Dinis-Ribeiro, Mário, Figueiredo, Céu, Keevil, Charles W., Vieira, Maria J. and Azevedo, Nuno F. (2013) Validation of a PNA-FISH method for the detection of Helicobacter pylori clarithromycin resistance in gastric biopsies. Journal of Clinical Microbiology. (doi:10.1128/JCM.00302-13). (PMID:23596234)

Record type: Article

Abstract

In here, we evaluated a previously established PNA-FISH method as a new diagnostic test for H. pylori clarithromycin resistance detection in paraffin embedded gastric biopsies. Both a retrospective and a prospective cohort study were conducted to evaluate the specificity and sensitivity of a PNA-FISH method to determine H. pylori clarithromycin resistance. In the retrospective study (n=30 patients), full agreement between PNA-FISH and PCR-sequencing was observed. When compared to the reference method (culture followed by Etest®), specificity and sensitivity of PNA-FISH were 90.9% (95% confidence interval [CI], 57.1-99.5%) and 84.2% (95% CI, 59.5-95.8%), respectively. In the prospective cohort (n=93 patients), 21 cases were positive by culture. For the patients harboring clarithromycin-resistant H. pylori, the method showed a sensitivity of 80.0% (95% CI, 29.9-98.9%) and specificity of 93.8% (95% CI, 67.7-99.7%). These values are likely to be underestimated, as in some discrepant results patients were infected by more than one strain. PNA-FISH appears to be a simple, quick and an accurate method for detecting
H. pylori clarithromycin resistance in paraffin embedded biopsies. It is also the only one of the methods assessed here that allows direct and specific visualization of this microorganism within the biopsies, a characteristic that allowed observing that different H. pylori strains can subsist very closely in the stomach.

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e-pub ahead of print date: 17 April 2013
Organisations: Centre for Biological Sciences

Identifiers

Local EPrints ID: 351707
URI: https://eprints.soton.ac.uk/id/eprint/351707
ISSN: 0095-1137
PURE UUID: f730beb7-0e91-4c96-ad90-6f8cb1e664ab
ORCID for Charles W. Keevil: ORCID iD orcid.org/0000-0003-1917-7706

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Date deposited: 30 Apr 2013 11:06
Last modified: 15 Aug 2019 00:47

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Contributors

Author: Laura Cerqueira
Author: Ricardo M. Fernandes
Author: Rui M. Ferreira
Author: Mónica Oleastro
Author: Fátima Carneiro
Author: Catarina Brandao
Author: Pedro Pimentel-Nunes
Author: Mário Dinis-Ribeiro
Author: Céu Figueiredo
Author: Maria J. Vieira
Author: Nuno F. Azevedo

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