Analysis of clinical isolates of Helicobacter pylori in Pakistan reveals high degrees of pathogenicity and high frequencies of antibiotic resistance.
BACKGROUND
Antibiotic resistance in Helicobacter pylori contributes to failure in eradicating the infection and is most often due to point and missense mutations in a few key genes.


METHODS
The antibiotic susceptibility profiles of H. pylori isolates from 46 Pakistani patients were determined by Etest. Resistance and pathogenicity genes were amplified, and sequences were analyzed to determine the presence of mutations.


RESULTS
A high percentage of isolates (73.9%) were resistant to metronidazole (MTZ), with considerable resistance to clarithromycin (CLR; 47.8%) and amoxicillin (AML; 54.3%) also observed. Relatively few isolates were resistant to tetracycline (TET; 4.3%) or to ciprofloxacin (CIP; 13%). However, most isolates (n = 43) exhibited resistance to one or more antibiotics. MTZ-resistant isolates contained missense mutations in oxygen-independent NADPH nitroreductase (RdxA; 8 mutations found) and NADH flavin oxidoreductase (FrxA; 4 mutations found). In the 23S rRNA gene, responsible for CLR resistance, a new point mutation (A2181G) and 4 previously reported mutations were identified. Pathogenicity genes cagA, dupA, and vacA s1a/m1 were detected frequently in isolates which were also found to be resistant to MTZ, CLR, and AML. A high percentage of CagA and VacA seropositivity was also observed in these patients. Phylogenetic analysis of partial sequences showed uniform distribution of the 3' region of cagA throughout the tree.


CONCLUSIONS
We have identified H. pylori isolates in Pakistan which harbor pathogenicity genes and worrying antibiotic resistance profiles as a result of having acquired multiple point and missense mutations. H. pylori eradication regimens should therefore be reevaluated in this setting.
Department of Gastroenterology, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3GE, UK; Department of Microbiology, Quaid-i-Azam University, Islamabad, 45320, Pakistan.
Faisal
F
Rasheed
Barry James
BJ
Campbell
Hanafiah
H
Alfizah
Andrea
A
Varro
Rabaab
R
Zahra
Yoshio
Y
Yamaoka
David Mark
DM
Pritchard
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D017291:Clarithromycin / Q000494:pharmacology; D015331:Cohort Studies; D024881:Drug Resistance, Bacterial; D005260:Female; D016481:Helicobacter Infections / Q000453:epidemiology* / Q000382:microbiology; D016480:Helicobacter pylori / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D006801:Humans; D008297:Male; D008795:Metronidazole / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D008875:Middle Aged; D008969:Molecular Sequence Data; D009154:Mutation; D010154:Pakistan / Q000453:epidemiology; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D014774:Virulence; D055815:Young Adult
D001426:Bacterial Proteins; D008795:Metronidazole; D017291:Clarithromycin
23S rRNA gene; Helicobacter pylori infection; antibiotic resistance; cagA; clarithromycin resistance; metronidazole resistance