Discrimination of epidemic and sporadic isolates of Acinetobacter baumannii by repetitive element PCR-mediated DNA fingerprinting (original) (raw)
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Journal of clinical microbiology, 1993
A polymerase chain reaction (PCR) technique was applied to the fingerprinting of different strains of Acinetobacter baumannii from a cluster of patients infected or colonized with the incriminated pathogen. The DNA was extracted by boiling and was subjected to PCR amplification by using the core sequence of the M13 phase as a single primer. The amplified products were separated by agarose gel electrophoresis and were detected by staining with ethidium bromide. In 1990, 49 multiresistant A. baumannii strains were isolated from 13 patients from the same intensive care unit of the Charité Hospital; 45 of these outbreak isolates obtained from 12 patients showed the same PCR patterns, indicating an epidemiological relatedness of these strains. Four strains isolated from the same patient belonged to another genetic group, as revealed by a distinct amplification pattern. Another single subtype of A. baumannii was identified as the causative agent in patients during a second outbreak at a d...
Scandinavian Journal of Infectious Diseases, 1996
An epidemiological investigation of an outbreak o f Acinetobacter baumannii among patients on 2 closely related intensive care units (ICU) was performed by molecular typing with interrepeat polymerase chain reaction (interrepent PCR). 31 A, baumannii isolates obtained from 15 ICU patients were characterized. All patients were infected or colonized with A. baumannii. After identification of the outbreak, 6 environmental isolates were collected from tap-water, sinks and cleaning detergents. PCR fingerprinting identified 3 genotypes among the outbreak-related strains. One predominant genotype was demonstrated in 14/15 patients and this genotype was also found among all environmental isolates. The cluster o f A. baumannii represented an outbreak of 1 genotype, suggesting cross-contamination. The finding of the identical genotype among all environmental strains indicated a common environmental source causing the outbreak. The outbreak was controlled after reimplementation of an effective disinfection of workplace surfaces. This survey proved interrepeat PCR to be a rapid and reliable method to differentiate A. baumannii strains, thereby allowing epidemiological surveillance of large amounts of strains and early interventions to control outbreaks.
Efficacy of two DNA fingerprinting methods for typing Acinetobacter baumannii isolates
Diagnostic Microbiology and Infectious Disease, 2001
Performance of macrorestriction and repetitive extragenic palindromic DNA sequence-based PCR (REP-PCR) to type Acinetobacter baumannii isolates was quantitatively estimated using a test population of 54 outbreak-related, 29 endemic infection-related and 17 epidemiologically-unrelated isolates. Reproducibility and stability for macrorestriction were 100%, and REP-PCR showed only slightly lower stability. Macrorestriction resolved 18 fingerprints and REP-PCR 10 DNA patterns, forming eight and seven clusters at 75% of similarity level, respectively. Intercluster band variation was Ͼ7 bands for both methods. Although, all endemic isolates, except one, were concordantly grouped by both methods, macrorestriction distinguished a greater number of subtypes over one year study. For outbreaks, the epidemiologic concordance for both methods was 88%. The discriminatory index for macrorestriction and REP-PCR was 0.884 and 0.877, respectively. In conclusion, both methods showed similar efficacy as epidemiological markers, and by concordance, this study demonstrated that for REP-PCR typing, a Ն7 bands difference seemed an appropriate threshold to identify unrelated strains.
The Journal of Infection in Developing Countries, 2015
Introduction: Determination of microbial genetic relatedness is important for improving efficiency of infection control measures during hospital outbreaks. This study aimed to analyze the clonal relationships of clinical and environmental Acinetobacter baumannii strains isolated during an outbreak in the intensive care unit (ICU) of a secondary care hospital in Saudi Arabia. Methodology: Twelve clinical and fourteen environmental A. baumannii isolates identified during an outbreak in February 2013 in the 14-bed adult intensive care unit of a tertiary care hospital in Riyadh, Saudi Arabia, were studied. Bacterial identification and antimicrobial susceptibility testing were carried out using Microscan Walkaway 96 automated system. Determination of clonal diversity was investigated by repetitive-sequence-based polymerase chain reaction (rep-PCR) with the semi-automated DiversiLab system. Results: The majority of the clinical isolates were from endotracheal tube aspirates (n = 9), one ...
2008
Using a repetitive-sequence-based (rep)-PCR (DiversiLab), we have molecularly typed Acinetobacter nosocomial bloodstream isolates (Acinetobacter baumannii [n ؍ 187], Acinetobacter pittii [n ؍ 23], and Acinetobacter nosocomialis [n ؍ 61]) obtained from patients hospitalized in U.S. hospitals over a 10-year period (1995-2004) during a nationwide surveillance study (Surveillance and Control of Pathogens of Epidemiological Importance [SCOPE]). Patterns of A. baumannii rep-PCR were compared to those of previously identified international clonal lineages (ICs) and were further investigated by multilocus sequence typing (MLST) to compare the two typing methods. Forty-seven of the A. baumannii isolates clustered with the previously defined IC 2. ICs 1, 3, 6, and 7 were also detected. The remaining 81 isolates were unrelated to the described ICs. In contrast, A. pittii and A. nosocomialis isolates were more heterogeneous, as determined by rep-PCR. Our MLST results were in good correlation with the rep-PCR clusters. Our study confirms previous data indicating the predominance of a few major clonal A. baumannii lineages in the United States, particularly IC 2. The presence in the United States of A. baumannii ICs 1, 2, and 3 from as early as 1995 suggests that global dissemination of these lineages was an early event.
Phenotypic and genetic relationship of Acinetobacter Baumannii isolates
Prilozi / Makedonska akademija na naukite i umetnostite, Oddelenie za biološki i medicinski nauki = Contributions / Macedonian Academy of Sciences and Arts, Section of Biological and Medical Sciences, 2011
Acinetobacter continues to rise. One of the main reasons is the emergence of multi-resistant strains, which cause outbreaks of infection involving several patients in a ward, in the intensive care unit and in different areas of the hospital. Many outbreaks of its infection or colonization in surgical, neonatal and burn intensive care units have been reported, but the epidemiology of these infections remains unclear. Aim: To investigate the relationship among the isolates of Acinetobacter baumannii, comparing some of their phenotypic and genetic features. Material and Methods: A total of 20 Acinetobacter baumanni isolates were included in the study. 12 strains of Acinetobacter baumannii were obtained within a week in July 2010, from neonates hospitalized at the paediatric intensive care unit and on the neonatal ward. Three strains were isolated from neonates at the paediatric intensive care unit three months ago. All the Acinetobacter baumannii strains were isolated from tracheal aspirates obtained from neonates with infection of the lower respiratory tract. Five additional Acinetobacter baumannii strains were included in the study as controls. They were isolated from wound swabs taken from adult patients with wound infection, hospitalized at the University Traumatology Clinic. Susceptibility of the bacterial strains to 13 different antimicrobial agents was determined by the disk diffusion method (Kirby-Bauer). Additional testing of the susceptibility was performed by the VITEK 2 system. RAPD-PCR fingerprinting was carried out using the following primer (5' GAAACAGCTATGACCATG-3'). Results: All A. baumannii isolates were multi-drug resistant. Antibiotic susceptibility-testing by the disk-diffusion method and automated VITEK 2 system showed 3 and 2 antimicrobial susceptibility patterns, respectively. RAPD-PCR assay of A. bau-Trajkovska-Dokic E, et al. Contributions, Sec. Biol. Med. Sci., XXXII/2 (2011), 157-168 mannii strains revealed two different RAPD-fingerprints. All the strains of A. baumannii isolated within a week in July 2010 from tracheal aspirates taken from neonates in the paediatric intensive care unit and neonates in the paediatric ward revealed the same RAPD-fingerprint, as well as 3 strains of A. baumannii isolated from tracheal aspirates taken from neonates in the paediatric intensive care unit three months ago. 5 strains of A. baumannii isolated from wound swabs of patients hospitalized at the Traumatology Clinic revealed a different RAPD-fingerprint. Conclusion: All the strains of A. baumannii isolated from neonates in the paediatric intensive care unit and paediatric ward were multi-drug resistant. Investigating the resistance patterns in multi-resistant isolates of Acinetobacter is a useful method which can predict the strain relationship. This method could be completed by at least one molecular method, such as the RAPD-PCR technique, which has shown itself to be a convenient and more reliable in interpreting the strain relationship of the A. baumannii isolates. Good infection control procedures, including phenotypic and molecular typing of A. baumannii isolates, are essential for preventing outbreaks of multi-drug resistant A. baumannii infections in our hospitals.
Journal of Clinical Microbiology, 2006
Members of the genus Acinetobacter are ubiquitous in soil and water and are an important cause of nosocomial infections. A rapid method is needed to genotype Acinetobacter isolates to determine epidemiology and clonality during infectious outbreaks. Multilocus PCR followed by electrospray ionization mass spectrometry (PCR/ESI-MS) is a method that uses the amplicon base compositions to genotype bacterial species. In order to identify regions of the Acinetobacter genome useful for this method, we sequenced regions of six housekeeping genes (trpE, adk, efp, mutY, fumC, and ppa) from 267 isolates of Acinetobacter. Isolates were collected from infected and colonized soldiers and civilians involved in an outbreak in the military health care system associated with the conflict in Iraq, from previously characterized outbreaks in European hospitals, and from culture collections. Most of the isolates from the Iraqi conflict were Acinetobacter baumannii (189 of 216 isolates). Among these, 111 isolates had genotypes identical or very similar to those associated with well-characterized A. baumannii isolates from European hospitals. Twenty-seven isolates from the conflict were found to have genotypes representing different Acinetobacter species, including 8 representatives of Acinetobacter genomospecies 13TU and 13 representatives of Acinetobacter genomospecies 3. Analysis by the PCR/ESI-MS method using nine primer pairs targeting the most information-rich regions of the trpE, adk, mutY, fumC, and ppa genes distinguished 47 of the 48 A. baumannii genotypes identified by sequencing and identified at the species level at least 18 Acinetobacter species. Results obtained with our genotyping method were essentially in agreement with those obtained by pulse-field gel electrophoresis analysis. The PCR/ ESI-MS genotyping method required 4 h of analysis time to first answer with additional samples subsequently analyzed every 10 min. This rapid analysis allows tracking of transmission for the implementation of appropriate infection control measures on a time scale previously not achievable.
The International Arabic Journal of Antimicrobial Agents
A total of 12 Acinetobacter baumannii isolates have been recovered from hospitalized patients at two hospitals in Jordan over two different periods of time (2006 and 2008). Phenotypic and biochemical characterization with antibiotic susceptibility testing indicated that all isolates were belonging to A. baumannii. A high degree of conservation of both the 16S-23S rRNA gene intergenic spacer (ITS) length and the ITS sequence was observed among the isolates, and their identities were further confirmed by amplified ribosomal DNA gene restriction analysis (ARDRA). The application of ITS sequence-based identification and ARDRA provide promising tools for the elucidation of the clinical significance of genospecies identification of A. baumannii.
Journal of Clinical Microbiology, 2012
Using a repetitive-sequence-based (rep)-PCR (DiversiLab), we have molecularly typed Acinetobacter nosocomial bloodstream isolates (Acinetobacter baumannii [n ؍ 187], Acinetobacter pittii [n ؍ 23], and Acinetobacter nosocomialis [n ؍ 61]) obtained from patients hospitalized in U.S. hospitals over a 10-year period (1995-2004) during a nationwide surveillance study (Surveillance and Control of Pathogens of Epidemiological Importance [SCOPE]). Patterns of A. baumannii rep-PCR were compared to those of previously identified international clonal lineages (ICs) and were further investigated by multilocus sequence typing (MLST) to compare the two typing methods. Forty-seven of the A. baumannii isolates clustered with the previously defined IC 2. ICs 1, 3, 6, and 7 were also detected. The remaining 81 isolates were unrelated to the described ICs. In contrast, A. pittii and A. nosocomialis isolates were more heterogeneous, as determined by rep-PCR. Our MLST results were in good correlation with the rep-PCR clusters. Our study confirms previous data indicating the predominance of a few major clonal A. baumannii lineages in the United States, particularly IC 2. The presence in the United States of A. baumannii ICs 1, 2, and 3 from as early as 1995 suggests that global dissemination of these lineages was an early event.
Polish journal of microbiology / Polskie Towarzystwo Mikrobiologów = The Polish Society of Microbiologists, 2008
In search of an effective DNA typing technique for Acinetobacter baumannii strains for hospital epidemiology use, the performance and convenience of a new target sequence was evaluated. Using known genomic sequences of Acinetobacter baumannii strains AR 319754 and ATCC 17978, we developed single-locus variable-number direct-repeat analysis using polymerase chain reaction-restriction fragment length polymorphism (DR-PCR/RFLP) method. A total of 90 Acinetobacter baumannii strains isolated from patients of the Clinical Hospital in Bydgoszcz, Poland, were examined. Initially, all strains were typed using macrorestriction analysis of the chromosomal DNA by pulsed-field gel electrophoresis (REA-PFGE). Digestion of the chromosomal DNA with the ApaI endonuclease and separation of the fragments by PFGE revealed 21 unique types. Application of DR-PCR/RFLP resulted in recognition of 12 clusters. The results showed that the DR-PCR/RFLP method is less discriminatory than REA-PFGE, however, the n...