Detection of Helicobacter pylori resistance to clarithromycin and fluoroquinolones in Brazil: A national survey.
OBJECTIVE
To evaluate bacterial resistance to clarithromycin and fluoroquinolones in Brazil using molecular methods.


METHODS
The primary antibiotic resistance rates of Helicobacter pylori (H. pylori) were determined from November 2012 to March 2015 in the Southern, South-Eastern, Northern, North-Eastern, and Central-Western regions of Brazil. Four hundred ninety H. pylori patients [66% female, mean age 43 years (range: 18-79)] who had never been previously treated for this infection were enrolled. All patients underwent gastroscopy with antrum and corpus biopsies and molecular testing using GenoType HelicoDR (Hain Life Science, Germany). This test was performed to detect the presence of H. pylori and to identify point mutations in the genes responsible for clarithromycin and fluoroquinolone resistance. The molecular procedure was divided into three steps: DNA extraction from the biopsies, multiplex amplification, and reverse hybridization.


RESULTS
Clarithromycin resistance was found in 83 (16.9%) patients, and fluoroquinolone resistance was found in 66 (13.5%) patients. There was no statistical difference in resistance to either clarithromycin or fluoroquinolones (P = 0.55 and P = 0.06, respectively) among the different regions of Brazil. Dual resistance to clarithromycin and fluoroquinolones was found in 4.3% (21/490) of patients. The A2147G mutation was present in 90.4% (75/83), A2146G in 16.9% (14/83) and A2146C in 3.6% (3/83) of clarithromycin-resistant patients. In 10.8% (9/83) of clarithromycin-resistant samples, more than 01 mutation in the 23S rRNA gene was noticed. In fluoroquinolone-resistant samples, 37.9% (25/66) showed mutations not specified by the GenoType HelicoDR test. D91N mutation was observed in 34.8% (23/66), D91G in 18.1% (12/66), N87K in 16.6% (11/66) and D91Y in 13.6% (9/66) of cases. Among fluoroquinolone-resistant samples, 37.9% (25/66) showed mutations not specified by the GenoType HelicoDR test.


CONCLUSIONS
The H. pylori clarithromycin resistance rate in Brazil is at the borderline (15%-20%) for applying the standard triple therapy. The fluoroquinolone resistance rate (13.5%) is equally concerning.
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Bruno Squarcio
BS
Sanches
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Gustavo Miranda
GM
Martins
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Karine
K
Lima
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Bianca
B
Cota
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Luciana Dias
LD
Moretzsohn
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Laercio Tenorio
LT
Ribeiro
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Helenice P
HP
Breyer
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Ismael
I
Maguilnik
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Aline Bessa
AB
Maia
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Joffre
J
Rezende-Filho
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Ana Carolina
AC
Meira
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Henrique
H
Pinto
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Edson
E
Alves
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Ramiro
R
Mascarenhas
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Raissa
R
Passos
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Julia Duarte
JD
de Souza
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Osmar Reni
OR
Trindade
Bruno Squarcio Sanches, Gustavo Miranda Martins, Karine Lima, Bianca Cota, Luciana Dias Moretzsohn, Ana Carolina Meira, Henrique Pinto, Edson Alves, Raissa Passos, Julia Duarte de Souza, Osmar Reni Trindade, Luiz Gonzaga Coelho, Instituto Alfa de Gastroenterologia, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais 30130-100, Brazil.
Luiz Gonzaga
LG
Coelho
D000293:Adolescent; D000328:Adult; D000368:Aged; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001706:Biopsy; D001938:Brazil; D017291:Clarithromycin / Q000494:pharmacology*; D024881:Drug Resistance, Bacterial; D004724:Endoscopy; D005260:Female; D024841:Fluoroquinolones / Q000494:pharmacology*; D016481:Helicobacter Infections / Q000382:microbiology; D016480:Helicobacter pylori / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008826:Microbial Sensitivity Tests; D008875:Middle Aged; D009154:Mutation; D013270:Stomach / Q000473:pathology; D055815:Young Adult
D000900:Anti-Bacterial Agents; D024841:Fluoroquinolones; D017291:Clarithromycin
Clarithromycin; Fluoroquinolones; Helicobacter pylori; Microbial drug resistance; Molecular diagnostic techniques
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