Isolation and Determination of Multidrug Resistant Pseudomonas Aeruginosa from Clinical Samples

Background: Pseudomonas aeruginosa is a clinically important pathogenic microbe in hospitalized patients. It is a major cause of mortality and morbidity having a number of mechanisms that make it antibiotic resistant. Considering the dearth of antimicrobial drugs to treat infection with this pathogen, it has become a necessity to open up new arena for treatment with this organism. Recently, there has been an up rise in the number of multidrug resistant pathogenic strains of Pseudomonas aeruginosa. Objective: Isolation and identification of multidrug resistant Pseudomonas aeruginosa from wound specimens and to evaluate the antibiotic resistant strains of this microbe. Methodology: One hundred and fifty clinical samples of wound were taken from hospitalized patients at Jinnah hospital Lahore during the period of October 2019 to April 2020. In total, twenty (20) isolates of Pseudomonas aeruginosa were identified using the cultural features, morphological characteristics and various biochemical tests plus the Vitek 2 system. Blue/green, brown /blue and yellow/green pigment production showed the presence and growth of Pseudomonas aeruginosa. Results: Percentage of Pseudomonas aeruginosa in females came out to be 15% as compared to 11.42% in males. This was followed by testing susceptibility of isolates of Pseudomonas aeruginosa to various antimicrobial drugs. Piperacillin/tazobactam and meropenem showed the highest efficacy against Pseudomonas aeruginosa. Highest resistance was exhibited against trimethoprim/sulfamethoxazole which was 75%. Conclusion: Most isolates showed multidrug resistance to four or more drugs. Development of multidrug resistance has emerged as a global problem with pathogens commonly causing infections becoming increasingly resistant to antimicrobial agents.

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A Comprehensive Evaluation of GeneLEAD VIII DNA Platform Combined to Deeplex Myc-TB® Assay to Detect in 8 Days Drug Resistance to 13 Antituberculous Drugs and Transmission of Mycobacterium tuberculosis Complex Directly From Clinical Samples

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2021.707244 ◽

2021 ◽

Vol 11 ◽

Author(s):

Isabelle Bonnet ◽

Vincent Enouf ◽

Florence Morel ◽

Vichita Ok ◽

Jérémy Jaffré ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Comprehensive Evaluation ◽

Drug Susceptibility ◽

Multidrug Resistant ◽

Drug Susceptibility Testing ◽

Clinical Samples ◽

Routine Practice ◽

Single Nucleotide Variants ◽

Sequencing Platform

The GeneLEAD VIII (Diagenode, Belgium) is a new, fully automated, sample-to-result precision instrument for the extraction of DNA and PCR detection of Mycobacterium tuberculosis complex (MTBC) directly from clinical samples. The Deeplex Myc-TB® assay (Genoscreen, France) is a diagnostic kit based on the deep sequencing of a 24-plexed amplicon mix allowing simultaneously the detection of resistance to 13 antituberculous (antiTB) drugs and the determination of spoligotype. We evaluated the performance of a strategy combining the both mentioned tools to detect directly from clinical samples, in 8 days, MTBC and its resistance to 13 antiTB drugs, and identify potential transmission of strains from patient-to-patient. Using this approach, we screened 112 clinical samples (65 smear-negative) and 94 MTBC cultured strains. The sensitivity and the specificity of the GeneLEAD/Deeplex Myc-TB approach for MTBC detection were 79.3% and 100%, respectively. One hundred forty successful Deeplex Myc-TB results were obtained for 46 clinical samples and 94 strains, a total of 85.4% of which had a Deeplex Myc-TB susceptibility and resistance prediction consistent with phenotypic drug susceptibility testing (DST). Importantly, the Deeplex Myc-TB assay was able to detect 100% of the multidrug-resistant (MDR) MTBC tested. The lowest concordance rates were for pyrazinamide, ethambutol, streptomycin, and ethionamide (84.5%, 81.5%, 73%, and 55%, respectively) for which the determination of susceptibility or resistance is generally difficult with current tools. One of the main difficulties of Deeplex Myc-TB is to interpret the non-synonymous uncharacterized variants that can represent up to 30% of the detected single nucleotide variants. We observed a good level of concordance between Deeplex Myc-TB-spoligotyping and MIRU-VNTR despite a lower discriminatory power for spoligotyping. The median time to obtain complete results from clinical samples was 8 days (IQR 7–13) provided a high-throughput NGS sequencing platform was available. Our results highlight that the GeneLEAD/Deeplex Myc-TB approach could be a breakthrough in rapid diagnosis of MDR TB in routine practice.

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Multidrug-resistant and extremely drug-resistant Pseudomonas aeruginosa in clinical samples from a tertiary healthcare facility in Nigeria

Turkish Journal of Pharmaceutical Sciences ◽

10.4274/tjps.galenos.2021.66066 ◽

2021 ◽

Vol 0 (0) ◽

pp. 0-0

Author(s):

Amaka Marian Awanye ◽

Chidozie Ngozi Ibezim ◽

Catherine Nonyelum Stanley ◽

Hannah Onah ◽

Iheanyi Omezurike Okonko ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Healthcare Facility ◽

Multidrug Resistant ◽

Clinical Samples ◽

Drug Resistant ◽

Tertiary Healthcare

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Phenotypic determination of phage susceptibility among multidrug-resistant bacteria isolated from clinical samples of patients of tertiary care center, Nepal

International Journal of Antimicrobial Agents ◽

10.1016/j.ijantimicag.2021.106421.15 ◽

2021 ◽

Vol 58 ◽

pp. 21002867

Author(s):

Sreska Srestha ◽

Sngita Sharma ◽

Junu Rai ◽

Poonam Yadav ◽

Nirnajan Shah ◽

...

Keyword(s):

Care Center ◽

Tertiary Care ◽

Multidrug Resistant ◽

Clinical Samples ◽

Resistant Bacteria ◽

Tertiary Care Center ◽

Multidrug Resistant Bacteria

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Rapid Detection of Carbapenemase Genes (Bla-Vim And Bla-Imp) By Multiplex Pcr in Gram-Negative Clinical Isolates

10.21203/rs.3.rs-111985/v1 ◽

2020 ◽

Author(s):

Mehraj Ansari ◽

Kulraj Rai ◽

Ganesh Rai ◽

Subhas Aryal ◽

Shiba Rai ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Multiplex Pcr ◽

Rapid Detection ◽

Carbapenem Resistance ◽

Multidrug Resistant ◽

Clinical Samples ◽

Resistant Bacteria ◽

Multidrug Resistant Bacteria ◽

Gram Negative ◽

Positive Results

Abstract BackgroundCarbapenems have been the choice of antibiotics for the treatment of infections caused by multidrug-resistant bacteria. However, during recent years, carbapenems resistant bacteria have emerged significantly. The main objective of this study was to determine the prevalence of carbapenemase (bla-VIM and bla-IMP) producing isolates among Enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter baumannii.ResultsOf the total 1,151 clinical samples, 253 (22.0%) showed growth positive. Of them, 226 (89.3%) were identified as members of Enterobacteriaceae, P. aeruginosa and A. baumannii. Among the 226 isolates, 106 (46.9%) were multidrug-resistant. Of the 106, 97 (91.5%) isolates showed resistance to at least one of the carbapenem used. Among the 97 isolates, 67 (69.1%) showed MHT positive results. bla-VIM and bla-IMP were detected in 40 and 38 isolates, respectively.ConclusionThis study determined the higher prevalence of MDR and carbapenem resistance among Enterobacteriaceae, P. aeruginosa and A. baumannii as detected by the presence of bla-VIM and bla-IMP genes.Keywords: Carbapenems, Carbapenemase, Modified Hodge Test, bla-VIM, bla-IMP

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Molecular characterization of metallo β-lactamase producing multidrug resistant Pseudomonas aeruginosa from various clinical samples

Indian Journal of Pathology and Microbiology ◽

10.4103/0377-4929.142670 ◽

2014 ◽

Vol 57 (4) ◽

pp. 579 ◽

Cited By ~ 10

Author(s):

Kalaivani Ramakrishnan ◽

Saranathan Rajagopalan ◽

Shashikala Nair ◽

Prashanth Kenchappa ◽

SheelaDevi Chandrakesan

Keyword(s):

Pseudomonas Aeruginosa ◽

Molecular Characterization ◽

Multidrug Resistant ◽

Clinical Samples

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Prevalence of Multidrug-resistant and Carbapenemase-producing Klebsiella Pneumoniae and Pseudomonas Aeruginosa Isolates in Tertiary Care Hospital in Kathmandu, Nepal

Nepal Medical College Journal ◽

10.3126/nmcj.v23i4.42209 ◽

2021 ◽

Vol 23 (4) ◽

pp. 290-296

Author(s):

Rojina Darnal ◽

Mehraj Ansari ◽

Ganesh Rai ◽

Kul Raj Rai ◽

Shiba Kumar Rai

Keyword(s):

Pseudomonas Aeruginosa ◽

Klebsiella Pneumoniae ◽

Tertiary Care Hospital ◽

Tertiary Care ◽

Multidrug Resistant ◽

Diffusion Method ◽

Clinical Samples ◽

Resistant Bacteria ◽

Care Hospital ◽

Disk Diffusion Method

Carbapenemases are the enzymes that catalyze β–lactam groups of antibiotics. The carbapenemase producers are resistant to β–lactam antibiotics and are usually multidrug-resistant bacteria challenging widely used therapeutics and treatment options. Therefore, the detection of carbapenemase activity among clinical isolates is of great therapeutic importance. We aimed to study the MDR and carbapenemase-producing Klebsiella pneumoniae and Pseudomonas aeruginosa isolated from various clinical samples at a tertiary care hospital in Nepal. A total of 3,579 clinical samples were collected from the patients visiting the Department of Microbiology, B&B Hospital, Gwarko, Lalitpur. The samples were processed to isolate K. pneumoniae and P. aeruginosa and then subjected to antibiotic susceptibility testing (AST) by the Kirby-Bauer disk diffusion method. Phenotypic detection of carbapenemase activity was performed in the imipenem-resistant isolates by the modified Hodge test (MHT). Of the total samples, 1,067 (29.8%) samples showed significant growth positivity, out of which 190 (17.3%) isolates were K. pneumoniae and 121 (11.3%) were P. aeruginosa. Multidrug resistance was seen in 70.5% of the K. pneumoniae isolates and 65.3% of the P. aeruginosa isolates. Carbapenemase production was confirmed in 11.9%, and 12.2% of the imipenem-resistant K. pneumoniae and P. aeruginosa isolates, respectively, by the MHT. This study determined the higher prevalence of MDR among K. pneumoniae and P. aeruginosa; however, carbapenemase production was relatively low.

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Diagnosis and Stratification of Pseudomonas aeruginosa Infected Patients by Immunochemical Quantitative Determination of Pyocyanin From Clinical Bacterial Isolates

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2021.786929 ◽

2021 ◽

Vol 11 ◽

Author(s):

Barbara Rodriguez-Urretavizcaya ◽

Nuria Pascual ◽

Carme Pastells ◽

Maria Teresa Martin-Gomez ◽

Lluïsa Vilaplana ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Limit Of Detection ◽

Release Kinetics ◽

Clinical Isolates ◽

Clinical Samples ◽

Mueller Hinton ◽

Highly Sensitive ◽

Kinetics Of ◽

Selection Of

The development of a highly sensitive, specific, and reliable immunochemical assay to detect pyocyanin (PYO), one of the most important virulence factors (VFs) of Pseudomonas aeruginosa, is here reported. The assay uses a high-affinity monoclonal antibody (mAb; C.9.1.9.1.1.2.2.) raised against 1-hydroxyphenazine (1-OHphz) hapten derivatives (PC1; a 1:1 mixture of 9-hydroxy- and 6-hydroxy-phenazine-2-carobxylic acids). Selective screening using PYO and 1-OHphz on several cloning cycles allowed the selection of a clone able to detect PYO at low concentration levels. The microplate-based ELISA developed is able to achieve a limit of detection (LoD) of 0.07 nM, which is much lower than the concentrations reported to be found in clinical samples (130 μM in sputa and 2.8 μM in ear secretions). The ELISA has allowed the investigation of the release kinetics of PYO and 1-OHphz (the main metabolite of PYO) of clinical isolates obtained from P. aeruginosa-infected patients and cultured in Mueller–Hinton medium. Significant differences have been found between clinical isolates obtained from patients with an acute or a chronic infection (~6,000 nM vs. ~8 nM of PYO content, respectively) corroborated by the analysis of PYO/1-OHphz levels released by 37 clinical isolates obtained from infected patients at different stages. In all cases, the levels of 1-OHphz were much lower than those of PYO (at the highest levels 6,000 nM vs. 300 nM for PYO vs. 1-OHphz, respectively). The results found point to a real potential of PYO as a biomarker of P. aeruginosa infection and the possibility to use such VF also as a biomarker for patient stratification[2] and for an effective management of these kinds of infections.

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Evaluation of blaGES-5 and bla veb-1 genes with multidrug-resistant extend, pandrug resistance patterns (MDR, XDR, PDR), and biofilm formation in Pseudomonas aeruginosa isolates

Cellular and Molecular Biology ◽

10.14715/cmb/2021.67.3.7 ◽

2021 ◽

Vol 67 (3) ◽

pp. 52-60

Author(s):

Fattma A. Ali ◽

Sevan Hassan Bakir ◽

Sayran Hamed Haji ◽

Bashdar M. Hussen

Keyword(s):

Pseudomonas Aeruginosa ◽

Biofilm Formation ◽

Phylogenetic Trees ◽

Antibiotic Sensitivity ◽

Multidrug Resistant ◽

Clinical Samples ◽

Sensitivity Testing ◽

Automated Method ◽

Resistance Patterns ◽

Vitek 2

Pseudomonas aeruginosa is a ubiquitous microorganism that is difficult to treat due to the increasing prevalence of multidrug resistance patterns. A total of 227 samples were taken from different clinical samples during the study period from January 2018 to December 2018. The isolates were identified with antibiotic sensitivity testing with ESBL by the Vitek-2 automated method. MDR, XDR, and PDR were determined. 40 (17.6%) isolates were positive for P. aeruginosa, maximum of ESBL and MBL. Positive isolates were detected in the burn, coexisting ESBL + MBL enzymes in 21 (52.5%) of our isolates. Imipenem followed by Meropenem were found to be effective against ESBL and MBL producers. Resistance was reached between 72-100% to 5 antibiotics. The frequency of PDR, MDR, and XDR were 5%, 50%, and 45%, respectively. The frequency of co-production between MDR, XDR, and PDR with MBL, ESBL, and Biofilm was 35%, 12.5% and 5%, respectively. Among the ESBLs, the frequency of distribution of bla VEB-1gene and blaGES-5 gene was 50% and 40 %, respectively. Bacterial isolates simultaneously carrying blaVEB-1 gene with multiple ?-lactamases of different classes of biofilm, MDR, PDR, and XDR as same as a coexisting blaGES-5 gene. One isolate was detected as new isolates registered in global gene bank as locally P. aeruginosa isolates in Erbil city (LOCUS MN900953). The phylogenetic trees of the blaVEB gene isolates were demonstrated a genotype closely related to others, deposited in GenBank similar to the P. aeruginosa gene; gene sequencing revealed a 99% similarity with other isolates deposited in GenBank.

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