Antibiotic resistance and clonal relatedness of Helicobacter pylori strains isolated from stomach biopsy specimens in northeast of Iran.

BACKGROUND: Resistant Helicobacter pylori to commonly used antimicrobial agents are associated with severe upper gastrointestinal disorders. To provide an epidemiological picture of H pylori and characterize the resistance pattern and genetic variation of clinical isolates, stomach biopsies from patients with functional dyspepsia were evaluated in northeast of Iran. MATERIALS AND METHODS: In this study, 80 patients were recruited. Finally, fifty H pylori strains were isolated from antrum and corpus biopsies by culturing on Columbia agar. All strains were identified by standard laboratory procedures. Susceptibility testing of antibiotics was performed using minimum inhibitory concentration test. Allele-specific primer (ASP)-PCR of 23S rRNA which associated with clarithromycin resistance was done among resistant strains. Moreover, cagA gene and polymorphism in vacA were detected. Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) was applied to investigate the genetic variations among all strains. RESULTS: Antibiotic resistance pattern of H pylori strains was as follows: 68% (34/50) to metronidazole, 50% (25/50) to rifampicin, 30% (15/50) to amoxicillin, 28% (14/50) to levofloxacin, 22% (11/50) to clarithromycin, and 16% (8/50) to tetracycline. Multidrug-resistant strains were observed in 19 strains (38%). ASP-PCR of 23S rRNA showed four strains had A2143G mutation, six strains had A2142G mutation, and one strain had a Wt+A2143G mutation. Amplification of virulence-associated genes revealed that cagA was present in 27 isolates (54%) and vacA in 36 isolates (72%). The most common genotype of H pylori was vacA s1am2 (40%) followed by vacA s2m2 (14%), vacA s1am1 (12%), vacA s1bm1 (4%), and vacA s1bm2 (2%). DNA fingerprinting pattern indicated a high heterogeneity among isolated strains. CONCLUSION: An alarming level of resistance to metronidazole and rifampicin and high heterogeneity among H pylori isolates highlighted the importance of continued monitoring of antimicrobial susceptibility and epidemiological surveillance of this pathogen.

Outbreak of hypervirulent Klebsiella pneumoniae harbouring blaVIM-2 among mechanically-ventilated drug-poisoning patients with high mortality rate in Iran.

OBJECTIVES: Carbapenem-resistant Klebsiella pneumoniae infections are associated with increased rates of treatment failure and death. Several studies have reported isolates with a combined hypervirulent and antimicrobial-resistant phenotype. METHODS: In this study, the molecular characteristics of hypervirulent K. pneumoniae (hvKP) isolated from mechanically-ventilated patients admitted to a toxicological intensive care unit (ICU) in Tehran, Iran, were examined. String test, antimicrobial susceptibility testing, virulence factors analysis and plasmid replicon typing were performed. The clonal relatedness of the isolates was analysed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). RESULTS: hvKP accounted for 9.4% (5/53) of K. pneumoniae isolated from ventilator-associated pneumonia among patients admitted to the ICU with acute drug poisoning. The mortality rate was 7.5% (4/53) among K. pneumoniae-infected patients. All fatal K. pneumoniae were hvKP isolates, were resistant to imipenem and harboured an aacA7, blaVIM-2 and dhfrI cassette arrangement in a class 1 integron. The isolates were shown to be ST23 (Pasteur scheme) and exhibited similar PFGE patterns. Plasmid analysis revealed a class 1 integron harbouring blaVIM-2 located on an ca. 45-kb IncN plasmid. CONCLUSION: Here we describe the emergence of VIM-2-producing hvKP serotype K1/ST23 in an outbreak with high mortality in a hospital toxicological ICU. It appears that we must alert and prepare the hospital's surveillance system for the appearance, expansion and clinical importance of new K. pneumoniae clones associated with high antimicrobial resistance and robust virulence capabilities.