scholarly journals Molecular Detection of Some Capsulargenes of Klebsiella pneumoniaeIsolated from Clinical Samples

2021 ◽  
Vol 26 (4) ◽  
pp. 138-145
Author(s):  
Amina A. Raheem ◽  
Ghaidaa J. Mohammed
Keyword(s):  
Klebsiella Pneumoniae ◽  
Virulence Factors ◽  
Virulence Factor ◽  
Molecular Detection ◽  
Opportunistic Pathogen ◽  
The Other ◽  
Clinical Samples ◽  
Biochemical Tests ◽  
Important Virulence Factor ◽  
Polymerase Chain

Klebsiella pneumoniae is an opportunistic pathogen that has been implicated as one of commonest cause of hospital and community acquired infections.The capsule of Klebsiella pneumoniae is an important virulence factor, involved in pathogenic mechanisms.So, this study aimed to isolateKlebsiella pneumoniae from different clinical samples from patients in Al-Diwaniyah teaching hospital and determine some virulence factors (capsulegenes)that used for serotyping of isolates. The study extended fromAugust  to November 2020.A total of 31 isolates from 80 different clinical samples identified as Klebsiella pneumoniae by traditional biochemical tests ,Vitek system and 16SrRNA gene.The existenceof three genes from7tested capsulargenes wasdetectedby Polymerase chain reaction.The commonness serotype were k2,k54,k57 where K2 detected in 4 (12.9 %), K54 in 12 (38.7 %),k57 in 4 (12.9).But, the other capsular polysaccaride genes k1,k3,k5,k20not detectedin all isolates of this study.

scholarly journals MOLECULAR DETECTION OF PROTEIN ENCODING GENES Virb11 ON BRUCELLA ABORTUS LOCAL ISOLATES ORIGIN OF PINRANG, NTT AND VACCINE STRAINS

2020 ◽  
Vol 21 (4) ◽  
pp. 503-511
Author(s):  
Maria Gladis Bupu Maze ◽  
Didik Handijatno ◽  
Wiwik Tyasningsih ◽  
Suwarno Suwarno ◽  
Agnes Theresia Soelih Estoepangestie ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Virulence Factor ◽  
Brucella Abortus ◽  
Molecular Detection ◽  
Livestock Population ◽  
Gram Staining ◽  
Protein Encoding ◽  
Important Virulence Factor ◽  
Pcr Method ◽  
Encoding Genes

Brucellosis in cattle is a disease caused by Brucella abortus due to the reduction in livestock population caused by abortion, stillbirth, weak birth, infertility and sterility. Brucella abortus has several potential virulence factors, i.e. virB11 gene that encodes VirB11 protein is an important virulence factor acts as an ATPase for assembling organelles when the bacteria replicate, helping to complete the bacterial cycle and agress to another cells. The aim of this study are to re-identification Brucella abortus and detect virB11 gene as encoding of B. abortus VirB11 protein in local isolates from Pinrang, NTT, strain vaccines S19 and RB51. The isolates Brucella abortus were re-cultured in Brucella agar base and re-identification is followed by microscopic with Gram staining and biochemical tested with urease, citrat, indol and TSIA test. virB11gene was detected with PCR method. The PCR result showed virB11 gene have DNA band 720 bp. virB11 gene are present in local isolates from Pinrang, NTT, strain vaccines S19 and RB51.

scholarly journals Molecular detection of β-lactamase genes in Klebsiella pneumoniae and Escherichia coli isolated from different clinical sources

2022 ◽  
Vol 67 (4) ◽  
pp. 170-180
Author(s):  
Kamal Ismael Bakr ◽  
Sherko Muhammed Abdul-Rahman ◽  
Rebwar Muhammad Hamasalih
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Molecular Detection ◽  
Multidrug Resistant ◽  
Clinical Samples ◽  
Chain Reaction ◽  
E Coli ◽  
Recent Emergence ◽  
Polymerase Chain ◽  
Pcr Test

The rising occurrence of infections generated by Escherichia coli and Klebsiella pneumoniae that produce extended-spectrum β-lactamase (ESBL) is reason for concern. Due to the recent emergence of multidrug-resistant microorganisms that develop ESBL. The purpose of this work was to detect the ESBLs in clinical isolates of E. coli and K. pneumoniae. 118 samples of E. coli and 63 isolates of K. pneumoniae were collected from clinical samples. Polymerase chain reaction was used to detect β-lactamase genes (i.e., blaTEM, blaSHV, and blaCTX-M). Phenotypic detection revealed that 48.31% and 85.19% of E. coli and K. pneumoniae produced ESBLs, respectively. Whereas screening of ESBL genes in both bacteria employing a multiplex PCR test revealed that 24.58% of the ESBL-producing E. coli strains contained blaTEM, 50.85% contained blaSHV, and 32.2% contained blaCTX-M. Nevertheless, in K. pneumoniae, 40.74% blaTEM, 35.19% blaSHV, and 64.81% blaCTX-M genes were present. Antimicrobial resistance profiles of E. coli and K. pneumoniae isolates to twenty antibiotics were observed to vary significantly. Additionally, it was determined that the majority of E. coli and K. pneumoniae isolates were multidrug resistant (MDR). Additionally, 80.51% of E. coli isolates were resistant to the AMC antibiotic, while 0.00% were resistant to IPM and MEM. From the other hand, the resistant proportion of K. pneumoniae isolates was heterogeneous, ranging from 69.84% against CAZ to 0.00% against CIP and G antibiotics. The blaSHV gene was the most widespread among different forms of ESBLs in E. coli, but the most common gene in K. pneumoniae isolates was blaCTX-M (64.81%).

scholarly journals Detection of LytA Genes in Streptococcus pneumoniae Isolated from sputum pneumonia patients

Biomedika ◽  
2020 ◽  
Vol 13 (1) ◽  
pp. 23-30
Author(s):  
Mustika Sari Hutabarat ◽  
Firdaus Hamid ◽  
Irawaty Djaharuddin ◽  
Alfian Zainuddin ◽  
Rossana Agus ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
False Negative ◽  
Pneumococcal Infection ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Clinical Samples ◽  
Biochemical Tests ◽  
Negative Results ◽  
False Negative Results ◽  
Polymerase Chain

Streptococcus pneumoniae (pneumococcus) is a Gram-positive facultative anaerobic bacterium that is a major cause of morbidity and mortality worldwide. But the lack of reporting of disease by this bacterium in Indonesia, one of the causes is because the diagnosis of pneumococcal infection is often clinically not typical and conventional methods which are still the standard gold method often give false-negative results. So the purpose of this study was to evaluate the performance of culture and molecular diagnostic methods using the Polymerase Chain Reaction (PCR) technique in detecting Streptococcus pneumoniae in sputum clinical samples using the Autolysin (LytA) gene which is a virulence factor of this bacterium. 57 isolates from 60 samples were confirmed as Streptococcus sp through microscopic identification, culture, and biochemical tests. Then the sensitivity test with an optochin test of 9 (9%) compared the results descriptively with the PCR technique using the Autolysin A (LytA) gene which was obtained more sensitive by 15 (25%).

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 PRESENCE OF MOLLICUTES AND MYCOPLASMA BOVIS IN NASAL SWABS FROM CALVES AND IN MILK FROM COWS WITH CLINICAL MASTITIS

2021 ◽  
Vol 28 ◽  
pp. 1-9
Author(s):  
Brunna Mayla Vasconcelos Adorno ◽  
Anelise Salina ◽  
Sâmea Fernandes Joaquim ◽  
Felipe De Freitas Guimarães ◽  
Bruna Churocof Lopes ◽  
...  
Keyword(s):  
Respiratory Diseases ◽  
Nasal Swab ◽  
Molecular Detection ◽  
Opportunistic Pathogen ◽  
Clinical Mastitis ◽  
Mycoplasma Bovis ◽  
Chain Reaction ◽  
Milk Samples ◽  
Nasal Swabs ◽  
Polymerase Chain

Mycoplasma bovis is part of the bovine respiratory tract microbiota but is considered an opportunistic pathogen of extreme importance in respiratory diseases of calves. It causes to the herd several diseases such as mastitis, polyarthritis, pneumonia and endometritis. This pathogen is highly contagious and animals with mastitis are potential disseminators of infection to the herd since they release from 106 to 108 CFU per mL milk. Similarly, animals with pneumonia eliminate, through respiratory secretions, high microbial loads of the agent. The present study aimed to perform molecular detection of Mycoplasma bovis in 185 milk samples from cows with clinical mastitis, as well as in 50 nasal swab samples from healthy calves with or without signs of pneumonia and born from cows with mastitis, all belonging to four dairy farms in Paraná State, where cases of mastitis had beendiagnosed. DNA extraction from both secretions was carried out according to the thermolysis method. For polymerase chain reaction (PCR), generic primers were employed to amplify the Mollicutes DNA and positive samples were subjected to PCR with primersspecific for M. bovis. Positivity for M. bovis was 3.78% in milk samples, regardless of the farm, and 20% in nasal swabs.

scholarly journals Analysis of Cuorum Sensing-Dependent Virulence Factors and Drug Resistance in Pseudomonas Aeruginosa Strains

2021 ◽  
pp. 46-51
Author(s):  
José José de Jesús Alba-Romero ◽  
Pablo Ruiz-Flores ◽  
Graciela Castro-Escarpulli ◽  
Sandra Isabel Hernández-González ◽  
Aurora Martínez-Romero ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Drug Resistance ◽  
Antibiotic Resistance ◽  
Risk Factor ◽  
Virulence Factors ◽  
Virulence Factor ◽  
Protein Analysis ◽  
Hospitalized Patients ◽  
Clinical Samples ◽  
Beta Lactamase

The objective was to analyze the virulence factors dependent on Cuorum Sensing and drug resistance in strains of Pseudomonas aeruginosa. Virulence factors such as pyocyanin, beta-lactamase, biofilm, and antibiotic resistance were determined in 95 strains of P. aeruginosa isolated from hospitalized patients. Genus and species were identified by protein analysis by MALDI-TOF. 100% of the strains were resistant to at least one drug and the highest proportion was 32 strains resistant to 4 drugs and 5 resistant PAM strains. In the analysis of virulence factors, 98.8% produce at least one virulence factor and 48.9% are beta-lactamase producers. Therefore, it is concluded that P. aeruginosa strains isolated from clinical samples constitute a risk factor for hospitalized patients.

scholarly journals Antibiotic resistance, virulence-associated genes analysis and molecular typing of Klebsiella pneumoniae strains recovered from clinical samples

AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Amir Mirzaie ◽  
Reza Ranjbar
Keyword(s):  
Antibiotic Resistance ◽  
Klebsiella Pneumoniae ◽  
Molecular Typing ◽  
Efflux Pump ◽  
Opportunistic Pathogen ◽  
Relevant Information ◽  
Multidrug Resistant ◽  
Clinical Samples ◽  
Microbiological Methods ◽  
High Level

AbstractKlebsiella pneumoniae is a multidrug-resistant (MDR) opportunistic pathogen that causes nosocomial infections. Virulence analysis and molecular typing as powerful approaches can provide relevant information on K. pneumoniae infection. In the current study, antibiotic resistance, virulence-associated genes analysis, as well as molecular typing of K. pneumoniae strains were investigated. Out of 505 clinical samples collected from hospitalized patients, 100 K. pneumoniae strains were isolated by standard microbiological methods and subjected to the phenotypic and genotyping analysis. The highest prevalence of resistance was observed against ciprofloxacin (75%), trimethoprim–sulfamethoxazole (73%) and nitrofurantoin (68%). Virulence associated genes including entB, traT, ybts, magA, iucC, htrA and rmpA were found in 80%, 62%, 75%, 5%, 30%, 72% and 48%, of the isolates, respectively. The prevalence of biofilm-associated genes including mrkA, fimH, and mrkD were equally 88% for all tested isolates. Moreover, the efflux pump genes including AcrAB, TolC and mdtK were observed in 41 (41%), 33 (33%) and 26 (26%) of the strains respectively. A significant statistical association was observed between MDR strains and high expression of efflux pump and biofilm genes. The K. pneumoniae strains were differentiated into 11 different genetic patterns using the repetitive element sequence-based PCR (rep-PCR) technique. High prevalence of resistance, presence of various virulence factors, high level of efflux pump, and biofilm gene expression in diverse clones of K. pneumoniae strains pose an important health issue in clinical settings.

scholarly journals Prevalence of Quinolones Resistance Proteins Encoding Genes (qnr genes) and Co-Resistance with β-lactams among Klebsiella pneumoniae Isolates from Iraqi Patients

2020 ◽  
Vol 17 (2) ◽  
pp. 0406
Author(s):  
Mustafa Mustafa et al.
Keyword(s):  
Klebsiella Pneumoniae ◽  
Biochemical Properties ◽  
The Other ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Missense Mutations ◽  
High Association ◽  
Resistance Proteins ◽  
Major Threat ◽  
Encoding Genes

This study investigated the prevalence of quinolones resistance proteins encoding genes (qnr genes) and co-resistance for fluoroquinolones and β-lactams among clinical isolates of Klebsiella pneumoniae.  Out of 150 clinical samples, 50 isolates of K. pneumoniae were identified according to morphological and biochemical properties. These isolates were collected from different clinical samples, including 15 (30%) urine, 12 (24%) blood, 9 (18%) sputum, 9 (18%) wound, and 5 (10%) burn. The minimum inhibitory concentrations (MICs) assay revealed that 15 (30%) of isolates were resistant to ciprofloxacin (≥4µg/ml), 11 (22%) of isolates were resistant to levofloxacin (≥8 µg/ml), 21 (42%) of isolates were resistant to ertapenem (≥8 µg/ml), 18 (36%) of isolates were resistant to imipenem (4- ≥16µg/ml), 43 (86%) of isolates were resistant to ceftriaxone (4- ≥64 µg/ml), 42 (84%) of isolates were resistant to ceftazidime (16-64 µg/ml), and 40 (80%) of isolates were resistant to cefepime (4- ≥16µg/ml). The results revealed that all fluoroquinolone resistant K. pneumoniae isolates were resistant for β-lactams that used in this study. Genotypic detection of qnr genes revealed that qnrS and qnrB were found in 38 (76%) and 18 (36%) of K. pneumoniae isolates, respectively. On the other hand, qnrA, qnrC, and qnrD were not found among K. pneumoniae isolates. DNA sequencing of qnrB gene revealed that the presence of silent and missense mutations that may have led to increase the resistance values of MICs for ciprofloxacin and levofloxacin. These variants were registered in NCBI at the accession numbers LC373260 and LC381730. The phylogenetic tree of qnrB variants showed a significant deviation of these variants from K. pneumoniae species. The spread of qnr genes among clinical isolates of  K. pneumoniae and high association observed between resistance to fluoroquinolones and β-lactams have led to a major threat to public health through development of MDR K. pneumoniae.  

scholarly journals Study of some Virulence Factors for Clostridium perfringens isolated from Clinical Samples and Hospital Environment and showing their Sensitivity to Antibiotics

2020 ◽  
Vol 11 (02) ◽  
pp. 253-256
Author(s):  
Zainab I Tahseen ◽  
Muhsin H. Edham ◽  
Asma S. Karomi
Keyword(s):  
General Hospital ◽  
Virulence Factors ◽  
Clostridium Perfringens ◽  
High Resistance ◽  
High Sensitivity ◽  
Hospital Environment ◽  
Clinical Samples ◽  
Bacterial Isolates ◽  
Biochemical Tests ◽  
The City

The study included taking 100 samples from different clinical sources, including wounds and burns, and from the hospital environment, in Kirkuk General Hospital and Azadi Teaching Hospital in the city of Kirkuk for the period from November 2017 to August 2018. The results of isolation and diagnosis showed the growth of 30 isolates that are positive for Clostridium perfringens, distributed between 15 isolates 37.5% from burns, 11 isolates 27.5% from wounds, and 4 isolates 20% from the hospital environment. These isolates were diagnosed based on microscopical, cultural and biochemical tests, in addition to being diagnosed with the Api 20A system. The sensitivity of isolates was tested toward a number of types of antibiotics, and all bacterial isolates showed a high sensitivity 100% against imipenem. As for the sensitivity to vancomycin, amikacin, tetracycline was 96.66, 90, and 66.66% respectively. While, all isolates showed a high resistance to metronidazole and colistin 100%, some virulence factors of C. perfringens isolates have been studied , and showed that all isolates (%100) have the ability to produce hemolysin, lecithinase, capsule, and spore, while 70% of the isolates produced DNAase.

scholarly journals Virulence factors and beta-lactamase production among vancomycin-resistant Enterococcus faecalis isolated from clinical samples and hospital environment

2016 ◽  
Vol 5 (1) ◽  
pp. 1 ◽  
Author(s):  
Moses David ◽  
Kennedy Imonitie ◽  
Richard Osuntoyinbo ◽  
Adetunji Olawale
Keyword(s):  
Enterococcus Faecalis ◽  
Virulence Factors ◽  
Test Organism ◽  
Opportunistic Pathogen ◽  
Hospital Environment ◽  
Clinical Samples ◽  
Beta Lactamase ◽  
Vana Gene ◽  
Surgical Ward ◽  
Vancomycin Resistant

Enterococcus faecalis, though opportunistic pathogen has emerged as one of the leading nosocomial pathogens and has been implicated in different human infections. The severity of the infections caused by this organism is largely due to its complex pathogenic process. The objective of this study was to determine the carriage of virulence factors and vanA gene among the strains of vancomycin-resistant E. faecalis isolated from hospitals. Standard methods were used for isolation, antibiotic susceptibility and detection of virulence factors in the isolates. A total of one hundred and twenty three (123) samples were screened out of which 69 (45.70 %) yielded E. faecalis. The highest percentage of the isolates was recovered from the environment followed by the clinical samples. Children surgical ward had the highest occurrence of the test organism followed by male surgical ward. All the isolates were resistant to both amoxycillin/clavulanic acid and ceftazidime, while 98.55%, 89.86% and 53.62% were resistant to ampicillin, cefuroxime and gentamicin respectively. Only twenty seven (39.13%) of the isolates were resistant to the vancomycin. Among the vancomycin-resistant isolates, haemolysin had the highest occurrence (60.29%) followed by caseinase (55.88%). A total of 16 (59.26%) were beta-lactamase positive while 8 (29.63%) out of the isolates (vancomycin-resistant) were non-biofilm former while vanA genes was detected in 9 (33.33%) of the isolates. This study gives an insight to antibiotic resistant pattern of circulating Enterococcus faecalis and also the isolate showed varying patterns of virulent factors.

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