scholarly journals Molecular Investigation of hemolytic phospholipase C Encoding Genes in Clinical Isolates of Pseudomonas aeruginosa

2018 ◽  
Vol 9 (SPL1) ◽  
Author(s):  
Rasha Hadi Saleh ◽  
Habeeb S Naher ◽  
Mohammed AK Al-saadi
Keyword(s):  
Phospholipase C ◽  
Virulence Genes ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Burn Wound ◽  
Molecular Investigation ◽  
Genes Encoding ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Encoding Genes

This study is aimed to isolate P.aeruginosa from different clinical cases and to detect the prevalence of virulence genes encoding hemolytic phospholipase C(plcH)in these clinical isolates. In this study a total of 422 clinical samples including burn,wound,ear,urine,abscess and stool were aseptically taken from out- and inpatients who admitted into two hospitals in Hilla City (Teaching Al-Hilla Hospital and Babylon Hospital for Maternity and children during a period of three months. All samples were subjected to bacterial cultivation for the isolation of P.aeruginosa. The isolated P.aeruginosa was diagnosed depended on morphological,biochemical and molecular standard characteristics. Hemolytic phospholipase Cencoding genes(plcH) were detected by PCR and the amplification products were separated in 1% agarose gels containing ethidium bromide. Out of 422 samples,P.aeruginosa was isolated from 54 samples (12.8%). The distribution of these isolates were: 22 (55%) from burn samples,2; (50%) from diabitics foot samples,8 (14.8%) from wound samples, 8 (32%) from ear samples,3 (11%) from abscess samples, 7 (4%) from stool samples,4 (4%) from urine samples and 0 sputum samples. The genotypic properties of hemolytic phospholipase C (plcH )toxins was detected by polymerase chain reaction (PCR). The results of this study revealed that(plcH )gene found in 13/20 (65%)of isolates.

scholarly journals Antibiotic Resistance and Expression Of Resistance-Nodulation-Division Pump- and Outer Membrane Porin-Encoding Genes inAcinetobacterSpecies Isolated from Canadian Hospitals

2013 ◽  
Vol 24 (1) ◽  
pp. 17-21 ◽  
Author(s):  
Dinesh Fernando ◽  
George Zhanel ◽  
Ayush Kumar
Keyword(s):  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Outer Membrane ◽  
Polymerase Chain Reaction Method ◽  
Clinical Isolates ◽  
Chain Reaction ◽  
Genes Encoding ◽  
Resistance Nodulation Division ◽  
Polymerase Chain ◽  
Encoding Genes

BACKGROUND: Bacterial pathogens belonging to the genusAcinetobactercause serious infections in immunocompromised individuals that are very difficult to treat due to their extremely high resistance to many antibiotics.OBJECTIVE: To investigate the role of resistance-nodulation-division (RND) pumps and porins in the antibiotic resistance ofAcinetobacterspecies collected from Canadian hospitals.METHODS: Clinical isolates ofAcinetobacterspecies collected from Canadian hospitals were analyzed for the expression of genes encoding RND pumps (adeB,adeG,adeJ,AciBau_2746and AciBau_2436) and outer membrane porins (carO, 33 kDa porin andoprD) using quantitative reverse transcription (qRT) polymerase chain reaction. Species identification of the isolates was performed using a multiplex polymerase chain reaction method forgyrB.RESULTS: The expression of RND pump-encoding genes was widespread in the clinical isolates ofAcinetobacterspecies, with each of the isolates expressing at least one RND pump.adeGwas found to be overexpressed in all of the isolates, whileadeBwas found to be overexpressed in only two isolates. Among the porin-encoding genes, the expression ofcarOwas considerably downregulated among the majority of isolates.CONCLUSION: The present study was the first to analyze the expression of RND pump- and porin-encoding genes in the clinical isolates ofAcinetobacterspecies from Canadian hospitals. The overexpression of genes encoding RND pumps and the downregulation of genes encoding porins was common in clinical isolates ofAcinetobacterspecies from Canadian hospitals, with the AdeFGH pump being the most commonly expressed RND pump.

Distribution of Iron Uptake Systems Encoding Genes Among the Clinical Isolates of Escherichia coli Compared to Foodstuffs Isolates

2020 ◽  
Vol 20 (4) ◽  
pp. 517-522
Author(s):  
Hassan Mahmoudi ◽  
Hadi Hossainpour ◽  
Mohammad Moradi ◽  
Mohammad Yousef Alikhani
Keyword(s):  
Escherichia Coli ◽  
Iron Uptake ◽  
Food Poisoning ◽  
Food Samples ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
E Coli ◽  
Microbiological Methods ◽  
Polymerase Chain ◽  
Encoding Genes

Background: Bacteria require iron ions to grow and infect the host, which, by using iron uptake systems, acquire free iron from their host cell. Escherichia coli is one of the most important pathogens to cause food poisoning and clinical infections. The aim of this study was to assess the distribution of iron uptake systems encoding genes in clinical isolates of E.coli compared to food samples isolates. Materials and Methods: This investigation was conducted to determine the prevalence of E. coli isolated from various sources of food and clinical specimens. The E. coli isolates confirmed by the standard microbiological methods. The isolates were examined for the presence of iut A and iuc A genes by specific primers using the polymerase chain reaction technique. Results: A total of 100 and 50 isolates of E. coli were collected from clinical samples and foodstuffs, respectively. The prevalence of E. coli in the food and clinical samples was 33.33% and 64.10%, respectively. The frequency of iut A and iuc A genes in the food and clinical isolates were 76%-84% and 86% - 83%, respectively. Conclusion: Our results showed that the prevalence of E. coli isolates with iut A and iuc A genes was relatively higher compared to many previous studies. The existence of these genes in E. coli strains is likely to be related to pathogenicity in those strains, which requires further studies in the future.

scholarly journals Presence of β-lactamase encoding genes in clinical isolates of Y. enterocolitica bioserotype 1B/O8mm isolated in Poland in 2009

2018 ◽  
pp. 149-157
Author(s):  
Katarzyna Zacharczuk
Keyword(s):  
Sensu Stricto ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Broth Microdilution ◽  
Genetic Determinants ◽  
Intrinsic Resistance ◽  
Variable Activity ◽  
Genes Encoding ◽  
Encoding Genes ◽  
Mic Value

Introduction: The intrinsic resistance to β-lactam antibiotics of Y. enterocolitica is mainly due to activity of two type β - lactamases, BlaA and BlaB, which are encoding by genes located on chromosome, blaA and ampC, respectively. High-pathogenicity bioserotype 1B/O8 of Y. enterocolitica has been isolated from clinical samples since 2004 year in Poland. The study shown that clinical isolates of Y. enterocolitica 1B/O8 collected from 2004 to 2009 in Poland constituted the same sensu stricto strain. The aim of present study was to determine the occurrence of genes encoding β-lactamases BlaA and BlaB with the MIC value characterization for selected antibiotics in clinical isolates of Yersinia enterocolitica bioserotype 1B/O isolated in Poland in 2009. Methods: Ampicilin and ticarcillin minimum inhibitory concentrations (MICs) for 64 clinical Y. enterocolitica bioserotype 1B/O8 isolates were determined by Etest and broth microdilution respectively. The blaA i blaB genes were detected by PCR. Results: All investigated isolates were resistant to ticarcillin (MIC 16 – 128 mg/ml), whereas 9,5% to ampicilin (MIC 12 mg/l) according to EUCAST interpretive criteria. All isolates yielded the expected amplicons of blaA and ampC genes. Conclusions: Observed differences in the resistance to ampicillin among analyzed isolates belonging to the epidemic sensu stricto strain can indicate the variable activity of genetic determinants of resistance to β-lactam antibiotics of Y. enterocolitica bioserotype 1B/O8.

Analysis of genetic polymorphisms affecting the four phospholipase C (plc) genes in Mycobacterium tuberculosis complex clinical isolates

Microbiology ◽  
2004 ◽  
Vol 150 (4) ◽  
pp. 967-978 ◽  
Author(s):  
C. Viana-Niero ◽  
P. E. de Haas ◽  
D. van Soolingen ◽  
S. C. Leão
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Phospholipase C ◽  
Genetic Polymorphisms ◽  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Clinical Isolates ◽  
Significant Similarity ◽  
Genomic Deletions ◽  
Genes Encoding ◽  
Mycobacterium Africanum

The Mycobacterium tuberculosis genome contains four highly related genes which present significant similarity to Pseudomonas aeruginosa genes encoding phospholipase C enzymes. Three of these genes, plcA, plcB and plcC, are organized in tandem (locus plcABC). The fourth gene, plcD, is located in a different region. This study investigates variations in plcABC and plcD genes in clinical isolates of M. tuberculosis, Mycobacterium africanum and ‘Mycobacterium canettii’. Genetic polymorphisms were examined by PCR, Southern blot hybridization, sequence analysis and RT-PCR. Seven M. tuberculosis isolates contain insertions of IS6110 elements within plcA, plcC or plcD. In 19 of 25 M. tuberculosis isolates examined, genomic deletions were identified, resulting in loss of parts of genes or complete genes from the plcABC and/or plcD loci. Partial plcD deletion was observed in one M. africanum isolate. In each case, deletions were associated with the presence of a copy of the IS6110 element and in all occurrences IS6110 was transposed in the same orientation. A mechanism of deletion resulting from homologous recombination of two copies of IS6110 was recognized in a group of genetically related M. tuberculosis isolates. Five M. tuberculosis isolates presented major polymorphisms in the plcABC and plcD regions, along with loss of expression competence that affected all four plc genes. Phospholipase C is a well-known bacterial virulence factor. The precise role of phospholipase C in the pathogenicity of M. tuberculosis is unknown, but considering the potential importance that the plc genes may have in the virulence of the tubercle bacillus, the study of isolates cultured from patients with active tuberculosis bearing genetic variations affecting these genes may provide insights into the significance of phospholipase C enzymes for tuberculosis pathogenicity.

scholarly journals Molecular characterization of clinical carbapenem-resistant Enterobacterales from Qatar

2021 ◽  
Author(s):  
Fatma Ben Abid ◽  
Clement K. M. Tsui ◽  
Yohei Doi ◽  
Anand Deshmukh ◽  
Christi L. McElheny ◽  
...  
Keyword(s):  
Common Species ◽  
Clinical Samples ◽  
Whole Genome ◽  
E Coli ◽  
Carbapenem Resistant ◽  
Genes Encoding ◽  
Encoding Genes ◽  
Sequence Types ◽  
Common Sequence

AbstractOne hundred forty-nine carbapenem-resistant Enterobacterales from clinical samples obtained between April 2014 and November 2017 were subjected to whole genome sequencing and multi-locus sequence typing. Klebsiella pneumoniae (81, 54.4%) and Escherichia coli (38, 25.5%) were the most common species. Genes encoding metallo-β-lactamases were detected in 68 (45.8%) isolates, and OXA-48-like enzymes in 60 (40.3%). blaNDM-1 (45; 30.2%) and blaOXA-48 (29; 19.5%) were the most frequent. KPC-encoding genes were identified in 5 (3.6%) isolates. Most common sequence types were E. coli ST410 (8; 21.1%) and ST38 (7; 18.4%), and K. pneumoniae ST147 (13; 16%) and ST231 (7; 8.6%).

scholarly journals Spread of TEM, VIM, SHV, and CTX-Mβ-Lactamases in Imipenem-Resistant Gram-Negative Bacilli Isolated from Egyptian Hospitals

2016 ◽  
Vol 2016 ◽  
pp. 1-15 ◽  
Author(s):  
El sayed Hamdy Mohammed ◽  
Ahmed Elsadek Fakhr ◽  
Hanan Mohammed El sayed ◽  
Said abd Elmohsen Al Johery ◽  
Wesam Abdel Ghani Hassanein
Keyword(s):  
Direct Sequencing ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Nucleotide Polymorphism ◽  
Single Nucleotide ◽  
Gram Negative ◽  
Carbapenem Resistant ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Therapeutic Problem

Carbapenem-resistant Gram-negative bacilli resulting fromβ-lactamases have been reported to be an important cause of nosocomial infections and are a critical therapeutic problem worldwide. This study aimed to describe the prevalence of imipenem-resistant Gram-negative bacilli isolates and detection ofblaVIM,blaTEM,blaSHV,blaCTX-M-1, andblaCTX-M-9genes in these clinical isolates in Egyptian hospitals. The isolates were collected from various clinical samples, identified by conventional methods and confirmed by API 20E. Antibiotic susceptibility testing was determined by Kirby-Bauer technique and interpreted according to CLSI. Production ofblaVIM,blaTEM,blaSHV, andblaCTX-Mgenes was done by polymerase chain reaction (PCR). Direct sequencing from PCR products was subsequently carried out to identify and confirm theseβ-lactamases genes. Out of 65 isolates, (46.1%) Escherichia coli, (26.2%) Klebsiella pneumoniae, and (10.7%) Pseudomonas aeruginosa were identified as the commonest Gram-negative bacilli. 33(50.8%) were imipenem-resistant isolates. 22 isolates (66.7%) carriedblaVIM, 24(72.7%) hadblaTEM, and 5(15%) showedblaSHV, while 12(36%), 6(18.2%), and 0(0.00%) harboredblaCTX-M-1,blaCTX-M-9, andblaCTX-M-8/25, respectively. There is a high occurrence ofβ-lactamase genes in clinical isolates and sequence analysis of amplified genes showed differences between multiple SNPs (single nucleotide polymorphism) sites in the same gene among local isolates in relation to published sequences.

Phylogeny of Vibrio cholerae Isolates from Patients with Cholera Disease in Babylon Province

2019 ◽  
Vol 10 (04) ◽  
pp. 701-707
Author(s):  
Ilham Abbas Bunyan ◽  
Hussein T. Abdulabbas ◽  
Lamees A. Abdul-Lateef
Keyword(s):  
Vibrio Cholerae ◽  
High Rate ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Biochemical Tests ◽  
Diagnostic Kits ◽  
Stool Samples ◽  
Polymerase Chain ◽  
El Tor ◽  
Issr Pcr

In the present study, a total of 35 stool samples were collected in the Central Health Laboratory of Babylon Province from patients presenting invasive cholera disease. The period of collection was from November 2017 to December 2018. Identification of Vibrio cholerae species was carried out by conventional methods, biochemical tests, diagnostic kits and then confirmed by PCR-based assay targeting the ompW gene. The results of this study reported that O1 serogroups were the predominant serogroup among all clinical samples with a high rate of 94.3% (N = 33), while only two isolates of non-O1/non-O139 (NAG) (5.7%) were documented as a causative agent to cholera or cholera-like disease. The phylogenetic relationship among all 35 studied strains elucidated by using polymerase chain reaction (PCR)-based fingerprinting assay (ISSR-PCR). The results of this assay showed grouping of Inaba strains into different clusters indicating that these strains were genetically diverse. Furthermore, V. cholerae El Tor O1 Ogawa strain (OG1) was closely related to strains of Inaba serotype. In contrast, NAG strains (NAG1 and NAG2) were not genetically similar to any of Inaba or Ogawa strains indicating different clone origin.

scholarly journals Diverse Escherichia coli pathovars of phylogroups B2 and D isolated from animals in Tunisia

2017 ◽  
Vol 11 (07) ◽  
pp. 549-556 ◽  
Author(s):  
Hajer Kilani ◽  
Mohamed Salah Abbassi ◽  
Sana Ferjani ◽  
Rakia Ben Salem ◽  
Riadh Mansouri ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Virulence Genes ◽  
Chain Reaction ◽  
E Coli ◽  
Class 1 ◽  
Genes Encoding ◽  
Polymerase Chain ◽  
Relationship Of

Introduction: The virulent Escherichia coli strains responsible for extraintestinal infections were mainly belonged to B2 and D phylogroups. However, no past studies have determinate via the presence of virulence genes the frequency of E. coli pathovars recovered from animals housed in farms in Tunisia. The aims of this study were to investigate 26 E. coli isolated from healthy and diarrheic animals and to determinate via the presence of virulence genes the frequency of pathovars. Methodology: Twenty-six E. coli isolates of phylogroups B2 (n = 14), B22 (n = 9), B23 (n = 5), and D2 (n = 12) were characterized. Genes encoding virulence factors (fimH,eaeA,aggC,papC, papG allele III, hlyA, east1, cnf1, exhA,stx1, stx2, iutA, fyuA, ibeA,and ipaH), and antibiotic resistance as well as class 1 and 2 integrons were searched by polymerase chain reaction (PCR). The genetic relationship of isolates was done by PFGE. Results: According to the occurrence of specific genes the 26 isolates were classified as:9 EAEC, 2 EHEC, 4 UPEC, 3 EPEC/EHEC and 1 NTEC. Therefore, 2 Ex-PEC and 5 APEC were presented amongst our strains. Some isolates (12) were clonal and the remaining was unrelated. Conclusions: Higher diversity of pathovars which carried diverse combinations of virulence genes in healthy isolates. In addition, it seems that the infections were caused by different mechanisms.

LABORATORY ERRORS FOR IDENTIFICATION OF THE ESCHERICHIA COLI STRAINS OF THE SEROLOGICAL GROUPS O6 AND O25 AS ACUTE INTESTINAL INFECTIONS

2020 ◽  
Vol 65 (6) ◽  
pp. 368-374
Author(s):  
Mariia A. Makarova ◽  
Zoya N. Matveeva ◽  
E. V. Smirnova ◽  
L. I. Semchenkova ◽  
I. A. Derevianchenko ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Virulence Genes ◽  
Beta Lactams ◽  
E Coli ◽  
Laboratory Errors ◽  
Intestinal Infections ◽  
Human Pathology ◽  
Genes Encoding ◽  
Stool Samples

Were studied the genes encoding the virulence factors of 221 strains: E. coli O6:H1 (194) and E. coli O25:H4 (27), isolated in 2014-2018 from stool samples of children and adults examined according to epidemic indications. Molecular methods included PCR with hybridization-fluorescence and electrophoresis detection of amplified products. The strains did not have virulence genes for diarrheagenic E. coli (DEC) pathogroups EPEC, ETEC, EIEC, EHEC, EAggEC, and belonged to the phylogenetic group B2. They contained from four to eight genes encoding virulence factors of ExPEC: E. coli O6:H1 - pap (68,6%), sfa (87,6%), fimH (96,4%), hly (62,4%), cnf (74,7%), iutA (97,9%), fyuA (95,9%), chu (100%); E. coli O25:H4 - pap (66,7%), afa (22,2%), fimH (100%), hly (44,4%), cnf (44,4%), iutA (100%) , fyuA (100%), chu (100%). The antimicrobial susceptibility testing to 6 classes of antimicrobials (beta-lactams, fluoroquinolones, aminoglycosides, nitrofurantoin, sulfanilamide, trimethoprim / sulfamethoxazole) according the EUCAST. 60,3% of E. coli O6:H1 were sensitive to antibiotics, E. coli O25:H4 remained sensitive to carbapenems and nitrofurans. Extended-spectrum cephalosporins resistance was due to the production ESBL (CTX-M). The 57,1% resistant strains of E. coli O6:H1 and 100% of E. coli O25:H4 strains belonged to the MDR phenotype. The XDR phenotype had one in five MDR strains of E. coli O6:H1 and E. coli O25:H4. All strains of E. coli O25:H4 belonged to ST131. Given the important role of E. coli in human pathology, detection of virulence genes should be performed to confirm the etiological significance of the isolated strain.

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