Molecular characterization of carbapenem-resistant strains of Klebsiella pneumoniae isolated from Iranian patients: first identification of blaKPC gene in Iran.

Multi-resistant Klebsiella pneumoniae has been considered a serious global threat. This study was initiated to investigate carbapenem resistance among K. pneumoniae isolates in Iran and to detect carbapenemases in resistant strains. From 2009 to 2012, 180 K. pneumoniae strains were collected from Tehran hospitals. Of the isolates, 42 isolates (23.3%) were resistant to meropenem, 29 isolates (16.1%) were resistant to ertapenem, and 14 isolates (7.7%) were resistant to imipenem. All of carbapenem-resistant isolates were also resistant to the third generation of cephalosporins. modified Hodge test was positive in 25 (59.5%) of carbapenem-resistant isolates showing carbapenemase production. bla(NDM) and bla(VIM) genes were identified in three and five carbapenem-resistant isolates, respectively. One isolate showed presence of bla(KPC) gene. Class 1 integrons were detected in 14 carbapenem-resistant isolates. The most important finding about class 1 integrons was identification of an integron containing metallo-ß-lactamase gene VIM-1 that also harbored dfrA27 and arr3 genes. It is important to note that K. pneumoniae carbapenemase and New Delhi metallo-beta-lactamase-positive isolates identified in this study showed resistance to the majority of routine antimicrobial agents, including all ß-lactams and other classes of antibiotics. To our knowledge, this is the first identification of bla(KPC) and bla(VIM-1) genes among isolates of K. pneumoniae in Iran.

Identification and characterization of class 1 integrons among atypical enteropathogenic Escherichia coli isolated from children under 5 years of age.

BACKGROUND AND OBJECTIVES: The therapeutic options for diseases caused by Escherichia coli are limited. In this study we investigated the presence of virulence factors among Enteropathogenic Escherichia coli (EPEC) strains and their antibiotic resistance patterns. The isolates were also checked for the presence of class1 integrons and gene cassettes. MATERIALS AND METHODS: This study included 70 EPEC strains isolated from children. Antimicrobial resistance patterns were determined using diffusion methods. The broth microdilution methods was used to determine the minimum inhibitory concentration. PCR was used to detect eaeA, bfpA genes. The 5' and 3' conserved sequences (CSs) of class 1 integrons and intI gene were amplified to investigate the presence of integrons and gene cassettes. RESULTS: Antimicrobial susceptibility testing showed that 4 (5.7%), 3 (4.2%), and 2 (2.8 %) isolates were resistant to ampicillin, trimethoprim-sulfamethoxazole, and ceftazidime, respectively. Resistance rates to ciprofloxacin and aztreonam were 1.4%. Thirteen (18.5%) isolates showed resistance to tetracycline, and 4 (5.7%) were kanamycin resistant. Class I integron detected in 22 (31.4%) isolates. All the gene cassettes found in class I integrons corresponded to different variants of dfr and aadA genes. CONCLUSION: Prevalence of class I integrons in EPEC strains was high. Presence of aadA and dfr gene cassettes in integrons represents high distribution of resistance determinants in EPEC strains.

Identification of botulinum toxin type in clinical samples and foods in Iran.

BACKGROUND: Botulism is a serious neuroparalytic disease caused by toxins of Clostridium botulinum. Botulinum toxin is produced under anaerobic conditions and is one of the most dangerous toxin in the world. Rapid diagnosis of botulism is very essential for successful therapy. In this study, we reviewed data of cases of botulism in Iran from April 2004 through March 2010. MATHERIALS AND METHODS: From a total of 1140 samples of suspected botulism samples, 477 serum, 294 stool, 111 gastric secretions, and 258 food samples were collected from 21 provinces. These samples belonged to 432 distinct patients. All samples were tested for botulism by mouse bioassay, a gold standard method for detection of botulism. RESULTS: From 1140 received samples, 64 (5.6 %) positive samples of botulism were identified. Of these, 14 (21.8 %) cases had toxin type A, seven (11 %) cases had toxin type B, 22 (34.3 %) cases had toxin type E, and seven (11 %) cases had toxin type AB. The toxin type could not been identified in 14 (21.8 %) cases. The highest positive results were in Gilan, Tehran, Golestan, and Hamedan provinces. Seafoods and locally- made cheese were the most implicated foods in type E and type A botulism, respectively. CONCLUSION: Accurate and rapid diagnosis of botulism is very important because every case of botulism can be a public health emergency. During the study period, the median number of positive cases per year was 2.7 (range: one to18). Therefore, it is suggested that all clinicians are required to submit the collected samples from patients with botulism symptoms to the botulism reference laboratory for specific diagnosis and confirmation of botulism.

First report of New Delhi metallo-beta-lactamase-1-producing Klebsiella pneumoniae in Iran.

New Delhi metallo-beta-lactamase (NDM-1) is a novel metallo-beta-lactamase (MBL). Sporadic cases of NDM-1 positive strains have been reported from different countries, suggesting a widespread dissemination. The aim of this study was the detection of MBLs in Enterobacteriaceae isolated from patients in Tehran hospitals. After identification tests, the susceptibility to the antibiotics was done by Kirby-Bauer method and broth microdilution. Carbapenem-resistant isolates were tested for carbapenemase production using the modified Hodge test (MHT). Carbapenem-resistant strains screened for bla(KPC) gene and genes encoding MBLs. Twenty-three isolates (6.3%) were resistant to meropenem, eleven isolates (3%) were resistant to ertapenem, and four isolates (1.1%) were resistant to imipenem. MHT was positive in 11 (47.8%) of the carbapenem-resistant isolates. In March 2011, we detected a multiple drug-resistant Klebsiella pneumoniae isolate that was resistant to all tested antibiotics except colistin. PCR confirmed that this isolate contained bla(NDM-1), bla(TEM), bla(SHV), and bla(CTX-M). This is the first report on the detection of MBL NDM-1 in Iran. The rapid spread of NDM-1-positive bacteria proved to be a major challenge for the treatment and control of infectious diseases.

Serotyping of Streptococcus pneumoniae isolated from Tehran by Multiplex PCR: Are serotypes of clinical and carrier isolates identical?

BACKGROUND AND OBJECTIVES: Streptococcus pneumoniae is the most common cause of invasive infections among both young children and elderly people. Common serotypes causing invasive diseases and the emergence of carriers of Streptococcus pneumoniae in Iran is not yet known. Past-vaccine surveillance studies of serotype prevalence patterns in Iran are necessary to monitor the epidemiology of Streptococcus pneumoniae. Because of variation of pneumococcal serotypes in different geographical regions, in this study we evaluated common serotypes causing pneumococcal infections and healthy carrier children in Tehran by Multiplex PCR. MATERIALS AND METHODS: A total of 150 nasopharyngeal swabs were collected from healthy children in Tehran between December 2011 and August a2012, and 100 clinical samples were collected. Identification was performed by biochemical and molecular tests. Serotyping was done by multiplex PCR. We designed primers based on the sequences available for the routine capsular types and combined them into six multiplex PCR. RESULTS: From 150 nasopharyngeal swabs, 40 isolates of Streptococcus pneumoniae were identified after identification tests. Thirty six clinical isolates were also detected among clinical samples. Four serotypes (19A, 6, 3, 23F) of S. pneumoniae accounted for 55.7% of both sets of strains isolated from nasal carriage and clinical samples. Serotype 19A was the most common serotype among both groups. CONCLUSION: The multiplex PCR approach was successfully adapted to identify serotypes from more than 91% of the isolates tested. Among S. pneumoniae isolates in Tehran, the most prevalent serotypes were similar among carriage and invasive isolates. Continued monitoring of common serotypes of Streptococcus pneumoniae is essential for future vaccine formulation in Iran.