Molecular typing of Acinetobacter baumannii clinical strains by enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR)

Gene Reports ◽  
2020 ◽  
Vol 18 ◽  
pp. 100542
Author(s):  
Elham Zarifi ◽  
Mehran Ghazalibina ◽  
Shamsoddin Mansouri ◽  
Korosh Morshedi ◽  
Roya Pourmajed ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Acinetobacter Baumannii ◽  
Molecular Typing ◽  
Enterobacterial Repetitive Intergenic Consensus ◽  
Chain Reaction ◽  
Clinical Strains ◽  
Polymerase Chain

scholarly journals Molecular Typing of Klebsiella pneumoniae Clinical Isolates by Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction

2020 ◽  
Vol 2020 ◽  
pp. 1-5
Author(s):  
Parinaz Sedighi ◽  
Omid Zarei ◽  
Kiana Karimi ◽  
Mohammad Taheri ◽  
Pezhman Karami ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Polymerase Chain Reaction ◽  
Klebsiella Pneumoniae ◽  
Molecular Typing ◽  
Genetic Relatedness ◽  
Clinical Isolates ◽  
Enterobacterial Repetitive Intergenic Consensus ◽  
Chain Reaction ◽  
Microbiological Methods ◽  
Polymerase Chain

Aim. Klebsiella pneumoniae is one of the most important causes of nosocomial infections, including pneumonia, sepsis, and urinary tract infection. Enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) technique is a quick, reliable, and cost-effective method for molecular typing of Enterobacteriaceae family members. This study aimed to detect genetic relatedness among K. pneumoniae isolates from hospitals in Hamadan city, using ERIC-PCR technique. Materials and Methods. A total of 72 K. pneumoniae isolates were collected from patients admitted to Besat and Sina hospitals. After detection and confirmation of K. pneumonia isolates by chemical and conventional microbiological methods, DNAs were extracted after 24 hours of incubation at 37°C, using the boiling method. ERIC-PCR technique was carried out, and the ERIC patterns were analyzed by online data analysis service (inslico.ehu.es). ERIC profiles were compared using Dice method and clustered by UPGMA (unweighted pair group method with arithmetic mean) program. Also, the samples were evaluated by PCR method for the detection of aerobactin gene within their genome. Finding. The genetic relatedness among K. pneumoniae isolates was studied, and results established the genetic diversity of the clinical isolates by detecting 25 different ERIC types, including 14 common types and 11 unique types. Also, none of the isolates had aerobactin gene. Discussion. The results of this study showed high genetic diversity among K. pneumoniae strains, indicating the polyclonal distribution of K. pneumoniae isolates in Hamadan hospitals. This diversity causes problems for the treatment of infections due to the circulation of diverse K. pneumoniae clones, which possibly have different antimicrobial susceptibility patterns.

Diagnostic Value of Multiplex Polymerase Chain Reaction in Detection of Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from Sepsis in Pediatrics

2021 ◽  
Vol 15 ◽  
Author(s):  
Sara Galeb ◽  
Maysaa El Sayed Zaki ◽  
Raghdaa Shrief ◽  
Rasha Hassan ◽  
Mohamed Anies
Keyword(s):  
Polymerase Chain Reaction ◽  
Pseudomonas Aeruginosa ◽  
Acinetobacter Baumannii ◽  
Multiplex Pcr ◽  
Stenotrophomonas Maltophilia ◽  
Multiplex Polymerase Chain Reaction ◽  
Diagnostic Value ◽  
Blood Cultures ◽  
Chain Reaction ◽  
Polymerase Chain

Background: Proper identification of the causative organism in pediatric sepsis is crucial for early diagnosis and prevention of septic shock and organ failure. Objectives: The present study aimed to evaluate the multiplex Polymerase Chain Reaction (PCR) to detect Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from positive blood cultures for these pathogens isolated from children, with hospital-acquired sepsis compared to the conventional biochemical reactions for identification of these organisms. Methods: This study was a cross-sectional study performed on 100 isolates from pediatric blood cultures, including Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. The study also included 100 isolates of Escherichia coli as a negative control. All isolates were identified by API 20NE and the multiplex PCR, with primers specific to the 3 tested bacteria. Results: Multiplex PCR was positive in 96% of isolates, and 4 isolates had negative results. False positive results were reported with three E. coli strains. Multiplex PCR identified all the isolates of Acinetobacter baumannii, 29 isolates of Pseudomonas aeruginosa, and 27 isolates of Stenotrophomonas maltophilia. Compared to the biochemical identification, the diagnostic value of the multiplex PCR revealed 96.04% sensitivity, 96.9% specificity, 97.00%, positive predictive value, 96.00% negative predictive value, and 96.50% accuracy. Conclusion: The present study highlights the diagnostic value of multiplex PCR to identify Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from positive blood cultures. Multiplex PCR was sensitive, specific, and accurate. The accuracy differs according to the organisms, with 100% accuracy for Acinetobacter baumannii.

Typing of Haemophilus paragallinarum Strains by Using Enterobacterial Repetitive Intergenic Consensus-Based Polymerase Chain Reaction

10.1637/7137 ◽  
2004 ◽  
Vol 48 (4) ◽  
pp. 890-895 ◽  
Author(s):  
V. E. Soriano ◽  
G. Téllez ◽  
B. M. Hargis ◽  
L. Newberry ◽  
C. Salgado-Miranda ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Enterobacterial Repetitive Intergenic Consensus ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals The Inc FII Plasmid and its Contribution in the Transmission of blaNDM-1 and blaKPC-2 in Klebsiella pneumoniae in Egypt

Antibiotics ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 266 ◽  
Author(s):  
Eman Ramadan Mohamed ◽  
Mamdouh Yones Ali ◽  
Nancy G F M Waly ◽  
Hamada Mohamed Halby ◽  
Rehab Mahmoud Abd El-Baky
Keyword(s):  
Polymerase Chain Reaction ◽  
Klebsiella Pneumoniae ◽  
Molecular Typing ◽  
University Hospital ◽  
Chain Reaction ◽  
Great Problem ◽  
E Coli ◽  
String Test ◽  
Resistance Patterns ◽  
Polymerase Chain

The emergence of blaKPC-2 and blaNDM-1 producing Klebsiella pneumoniae represents a great problem in many Egyptian hospitals. One hundred and twenty-six K. pneumoniae isolates from patients admitted to Assiut University Hospital were identified by an API20E kit. Carbapenemase-producing K. pneumoniae (CPKP) was detected by the modified carbapenem inactivation method (mCIM), the EDTA-modified carbapenem inactivation method (eCIM), and an E-test. Based on the polymerase chain reaction, all isolates were negative for bla-VIM-1 and bla-IMP-1, fifteen of these isolates were positive for both blaKPC-2 and blaNDM-1, two isolates were positive for blaKPC-2 only, and twenty-eight isolates were positive for bla-NDM-1 only. Although one isolate was positive for the string test, all CPKP isolates were negative for capsular genes. Only 71.1% of CPKP transferred their plasmids to their corresponding transconjugants (E. coli J53). The resistance patterns of the clinical isolates and their transconjugates were similar, except for 12 isolates, which showed differences with their transconjugates in the resistance profile of four antibiotics. Molecular typing of the plasmids based on replicon typing showed that Inc FIIK and FII plasmids predominated in isolates and their transconjugants carrying blaKPC-2 and/or blaNDM-1. Conjugative Inc FII plasmids play an important role in the spread of CPKP, and their recognition is essential to limit their spread.

scholarly journals Molecular Epidemiology of Rhodococcus equi Based on traA, vapA, and vapB Virulence Plasmid Markers

2007 ◽  
Vol 196 (5) ◽  
pp. 763-769 ◽  
Author(s):  
Alain A Ocampo-Sosa ◽  
Deborah A Lewis ◽  
Jesús Navas ◽  
Frances Quigley ◽  
Raquel Callejo ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Typing ◽  
Rhodococcus Equi ◽  
Virulence Plasmid ◽  
Chain Reaction ◽  
Gene Markers ◽  
Typing System ◽  
New Type ◽  
Polymerase Chain ◽  
Related Plasmid

AbstractMolecular typing of the actinomycete Rhodococcus equi is insufficiently developed, and little is known about the epidemiology and transmission of this multihost pathogen. We report a simple, reliable polymerase chain reaction typing system for R. equi based on 3 plasmid gene markers: traA from the conserved conjugal transfer machinery and vapA and vapB, found in 2 different plasmid subpopulations. This “TRAVAP” typing scheme classifies R. equi into 4 categories: traA+/vapA+B−, traA+/vapA−B+, traA+/vapAB−, and traA−/vapAB− (plasmidless). A TRAVAP survey of 215 R. equi strains confirmed the strong link between vapA (traA+/vapA+B− plasmids) and horse isolates and revealed other host-related plasmid associations: between traA+/vapA−B+ and pigs and between traA+/vapAB−—a new type of R. equi plasmid—and cattle. Plasmidless strains were more frequent among isolates from nonpathological specimens. All plasmid categories were common in human isolates, which possibly reflects the predominantly opportunistic nature of R. equi infection in this host and a zoonotic origin.

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