Molecular resistance testing of Helicobacter pylori in gastric biopsies.
OBJECTIVE
To evaluate simultaneous diagnosis of infection and molecular resistance testing of Helicobacter pylori.


METHODS
Gastric biopsies were obtained from 26 rapid urease-positive and 51 rapid urease-negative test kits used to diagnose H pylori infection. Following glass bead-assisted DNA isolation, amplification of H pylori 16S ribosomal DNA (rDNA), glmM, and 23S rDNA target genes was performed.


RESULTS
Helicobacter pylori DNA was successfully amplified from 100% (26/26) of urease-positive and 3.9% (2/51) of urease-negative gastric biopsies. Subsequent restriction enzyme-mediated digestion of 23S rDNA amplification products revealed that 17% (4/24) of urease-positive and H pylori DNA-positive biopsy specimens contained point mutations (A2142G or A2143G) associated with clarithromycin resistance. Helicobacter pylori DNA from gastric biopsies was successfully amplified 8 weeks following rapid urease testing.


CONCLUSIONS
Helicobacter pylori genotyping may be used to detect macrolide-resistant H pylori in individuals prior to initiation of therapy or in patients refractory to anti-H pylori therapy. Two urease-negative specimens yielded Helicobacter DNA distinct from that of H pylori and indicated the need for further investigations of Helicobacter species present in the human stomach.
Department of Medical Laboratory Science, Northeastern University, Boston, MA, USA.
J A
JA
Peña
J G
JG
Fox
M J
MJ
Ferraro
J
J
Versalovic
D000900:Anti-Bacterial Agents / Q000627:therapeutic use; D001706:Biopsy; D017291:Clarithromycin / Q000627:therapeutic use; D017931:DNA Primers / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D004352:Drug Resistance, Microbial; D005753:Gastric Mucosa / Q000382:microbiology / Q000473:pathology*; D016481:Helicobacter Infections / Q000175:diagnosis* / Q000188:drug therapy*; D016480:Helicobacter pylori / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods*; D009154:Mutation; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D013272:Stomach Diseases / Q000201:enzymology / Q000382:microbiology / Q000473:pathology*; D014510:Urease / Q000378:metabolism
D000900:Anti-Bacterial Agents; D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D014510:Urease; D017291:Clarithromycin