Genotyping Approach Research Articles

Genome-scale analysis of Mycobacterium avium complex isolates from Portugal reveals extensive genetic diversity

Opportunist infections caused by nontuberculous mycobacteria (NTM) have emerged as a significant public health problem. Among these, species of the Mycobacterium avium complex (MAC) are the main responsible for the increase in the number of human disease cases. In order to address the current needs in the detection and surveillance of MAC disease cases, we evaluated different species classification methodologies (BLASTn-based marker-gene approach, Kraken v2, rMLST and MLST databases) and their congruence with a core-SNP phylogenetic approach, based on whole genome sequencing (WGS) data. For this purpose, we used a collection of 142 MAC isolates from Portuguese patients diagnosed between 2014 and 2022. The marker-gene approach (based on the rpoB, hsp65 and groEL genes), showed the best results, allowing the identification of the 142 MAC isolates to the species/subspecies level (M. avium subsp. hominissuis, M. intracellulare, M. intracellulare subsp. chimaera, M. intracellulare subsp. yongonense, M. marseillence and M. colombiense). Additionally, we performed drug susceptibility testing that confirmed clarithromycin efficacy as a first-line treatment for MAC disease, as 93 % of the Portuguese isolates were susceptible. Using a core-SNP approach we also performed an in-depth phylogenetic analysis within each identified species group, and despite the high genetic diversity within the MAC species, we were able to clearly distinguish all the species/subspecies and identify genetic clusters with epidemiological potential.We highlight not only the need for the standardization of an appropriate genotyping approach for species identification and management of MAC disease, but also a more robust large-scale WGS data analysis, in a One Health perspective, in order to identify potential routes of transmission.

Implementation of HLA-related genotype-guided prescribing in Singapore.

In an effort to expedite the publication of articles, AJHP is posting manuscripts online as soon as possible after acceptance. Accepted manuscripts have been peer-reviewed and copyedited, but are posted online before technical formatting and author proofing. These manuscripts are not the final version of record and will be replaced with the final article (formatted per AJHP style and proofed by the authors) at a later time. To describe the implementation of human leukocyte antigen (HLA)-related genotype-guided prescribing in Singapore. Various HLA alleles have been implicated in drug hypersensitivity syndromes (DHS). These include HLA-B*15:02, which has been associated with carbamazepine-induced Stevens-Johnson syndrome/toxic epidermal necrolysis, HLA-B*58:01, which has been associated with increased risk of severe cutaneous adverse reactions with allopurinol use, and HLA-B*57:01, which has been associated with increased risk of hypersensitivity reactions with abacavir use. Integrating pharmacogenomics into patient care through genotype-guided prescribing potentially optimizes use of these drugs by reducing DHS-related and healthcare costs. We describe the prevalence of HLA-related DHS in Singapore, the cost-effectiveness of genotype-guided prescribing, and local policies and guidelines, as well as the impact of genotype-guided prescribing where available. HLA-related genotype-guided prescribing has the potential to reduce the incidence of DHS and decrease healthcare costs, as seen in the success with carbamazepine. However, not all genotype-guided prescribing is cost-effective when implemented across the population, as was evident from local studies for allopurinol and abacavir. The cost-effectiveness of such measures may change over time with new data (eg, allele frequencies, test costs, drug prices, genotyping approach) and should be evaluated periodically and locally. Implementation of preemptive pharmacogenomics panel testing as part of routine clinical care may shift the threshold for cost-effectiveness and brings promise of further optimization of pharmacotherapy through precision medicine.

The role of salicylic acid in modulating phenotyping in spring wheat varieties for mitigating drought stress

Climate change-related droughts that recur frequently are one of the biggest obstacles to wheat (Triticum aestivum L.) productivity. Worldwide, attempts are being done to establish drought-resistant cultivars. However, progress is slow since drought tolerance is a complex trait controlled by numerous genes, and its expression is influenced by the environment. Phenotypic, biochemical physiological, and genotyping approaches are highlighted as critical research components for leveraging genetic variation in eight wheat genotypes. Treatments included eight spring wheat genotypes (IPK_040, IPK_046, IPK_050, IPK_071, IPK_105, WAS_007, WAS_024 and WAS_031), normal irrigation (NI), drought stress (D) (30% field capacity (FC)), normal irrigation with 0.5 mM SA (NSA), and drought treated with SA (DSA). The results revealed that there was a reduction in relative water content, an increase membrane leakage, and leaf chlorophyll content under drought stress. SA induced the defense responses against drought by increasing the osmolytes and the antioxidative enzymes activities. Compared to the NI group, the DSA treatment improved the water regulation, antioxidant capacity, and drought stress resistance. SA significantly reduced the deleterious effects of water stress on phenotyping more in WAS_ 024 and IPK_ 105 genotypes. The most responsive genotypes to salicylic acid were IPK_ 046 among the IPK genotypes, whereas WAS_031 genotype was amongst WAS genotypes based on the morpho-physiological traits. The findings of this study give a solid foundation for assessing drought resistance in T. aestivum and developing cultivation-specific water management methods.Graphical

A large structural variant collection in Holstein cattle and associated database for variant discovery, characterization, and application

BackgroundStructural variants (SVs) such as deletions, duplications, and insertions are known to contribute to phenotypic variation but remain challenging to identify and genotype. A more complete, accessible, and assessable collection of SVs will assist efforts to study SV function in cattle and to incorporate SV genotyping into animal evaluation.ResultsIn this work we produced a large and deeply characterized collection of SVs in Holstein cattle using two popular SV callers (Manta and Smoove) and publicly available Illumina whole-genome sequence (WGS) read sets from 310 samples (290 male, 20 female, mean 20X coverage). Manta and Smoove identified 31 K and 68 K SVs, respectively. In total the SVs cover 5% (Manta) and 6% (Smoove) of the reference genome, in contrast to the 1% impacted by SNPs and indels. SV genotypes from each caller were confirmed to accurately recapitulate animal relationships estimated using WGS SNP genotypes from the same dataset, with Manta genotypes outperforming Smoove, and deletions outperforming duplications. To support efforts to link the SVs to phenotypic variation, overlapping and tag SNPs were identified for each SV, using genotype sets extracted from the WGS results corresponding to two bovine SNP chips (BovineSNP50 and BovineHD). 9% (Manta) and 11% (Smoove) of the SVs were found to have overlapping BovineHD panel SNPs, while 21% (Manta) and 9% (Smoove) have BovineHD panel tag SNPs. A custom interactive database (https://svdb-dc.pslab.ca) containing the identified sequence variants with extensive annotations, gene feature information, and BAM file content for all SVs was created to enable the evaluation and prioritization of SVs for further study. Illustrative examples involving the genes POPDC3, ORM1, G2E3, FANCI, TFB1M, FOXC2, N4BP2, GSTA3, and COPA show how this resource can be used to find well-supported genic SVs, determine SV breakpoints, design genotyping approaches, and identify processed pseudogenes masquerading as deletions.ConclusionsThe resources developed through this study can be used to explore sequence variation in Holstein cattle and to develop strategies for studying SVs of interest. The lack of overlapping and tag SNPs from commonly used SNP chips for most of the SVs suggests that other genotyping approaches will be needed (for example direct genotyping) to understand their potential contributions to phenotype. The included SV genotype assessments point to challenges in characterizing SVs, especially duplications, using short-read data and support ongoing efforts to better characterize cattle genomes through long-read sequencing. Lastly, the identification of previously known functional SVs and additional CDS-overlapping SVs supports the phenotypic relevance of this dataset.

Comprehensive insight on multidrug resistance and virulence genes of ESBL-producing E. coli from different surface water sources in Bangladesh

ABSTRACT Environmental dissemination of multidrug resistance (MDR) has emerged as a global concern worldwide, particularly in developing countries like Bangladesh where the waste disposal system is inadequate. The present investigation was intended to comprehend the drug resistance and virulence pattern of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli (E. coli) in surface water samples of Bangladesh, which are adjoining poultry farms, agricultural fields, pharmaceutical companies, and textile industries. A total of 61 E. coli with ESBL production were identified from 16 water samples based on phenotypic and genotypic approaches. MDR was observed in 96% (n = 59) of the isolates with the multiple antibiotic resistance (MAR) index in the range of 0.3–0.9. The most prevalent MDR phenotype was non-susceptible to Ampicillin–Azithromycin–Nalidixic acid, with the highest percentage in the isolates from samples near agricultural fields. The frequencies of three ESBL resistance genes were detected as blaTEM (63%), blaCTX-M-13 (60%), and blaSHV (14%). Approximately 11% of the E. coli isolates were revealed as virulent gene positive, with the predominant one (eagg) specific for Enteroaggregative E. coli (EAEC). This study implies that the aquatic environment could be a potent means of exposure and transmission of bacterial antibiotic resistance and their genetic determinants in Bangladesh.

Multiplex PCR Approach for Rapid African Swine Fever Virus Genotyping.

African swine fever virus (ASFV) has been spreading through Europe, Asia, and the Caribbean after its introduction in Georgia in 2007 and, due to its particularly high mortality rate, poses a continuous threat to the pig industry. The golden standard to trace back the ASFV is whole genome sequencing, but it is a cost and time-intensive methodology. A more efficient way of tracing the virus is to amplify only specific genomic regions relevant for genotyping. This is mainly accomplished by amplifying single amplicons by PCR followed by Sanger sequencing. To reduce costs and processivity time, we evaluated a multiplex PCR based on the four primer sets routinely used for ASFV genotyping (B646L, E183L, B602L, and intergenic I73R-I329L), which was followed by Nanopore ligation-based amplicon sequencing. We show that with this protocol, we can genotype ASFV DNA originating from different biological matrices and correctly classify multiple genotypes and strains using a single PCR reaction. Further optimization of this method can be accomplished by adding or swapping the primer sets used for amplification based on the needs of a specific country or region, making it a versatile tool that can speed up the processing time and lower the costs of genotyping during ASFV outbreaks.

Molecular Marker Techniques and Genotypic Characterization Approaches in Plant Breeding

Plant breeding, often known as the science of plant development, is the study and practice of modifying a plant’s genetic makeup through a variety of breeding techniques to produce higher-quality, more prolific, and more resistant to harsh environmental circumstances. Classical breeding programs are indispensable techniques for increasing yields and improving plant characteristics, but they are progressing too slowly to meet the increasing food demand of the rapidly growing world population alone. Considering that the development periods of plants are generally long in traditional plant breeding, the opportunity to develop higher quality and more productive species that are more resistant to abiotic and biotic stress factors is very limited. Because there are multiple steps required in producing new plant varieties, including hybridization, selection, and testing, the process of creating a new variation takes several years. However, it is important to rapidly develop plant varieties with desirable characteristics to meet the increasing food demand of the rapidly growing world population, so the application of biotechnological methods integrated into plant breeding and combined with traditional methods can help reduce food shortages. Today, with the quick acceleration of biotechnology, molecular DNA marker technology has been developed in plant breeding and very important developments have been experienced. Thanks to the development of molecular tools for genetic research aimed at improving agricultural traits in plants related to crop yield, crop quality, or tolerance to adverse environmental conditions, we now have a much better understanding of plant genetics and the architecture and function of plant genomes. Therefore, it is of critical importance to revise current breeding procedures by incorporating molecular markers into breeding programs in the future.

Rapid Phenotypic and Genotypic Antimicrobial Susceptibility Testing Approaches for Use in the Clinical Laboratory.

The rapid rise in increasingly resistant bacteria has become a major threat to public health. Antimicrobial susceptibility testing (AST) is crucial in guiding appropriate therapeutic decisions and infection prevention practices for patient care. However, conventional culture-based AST methods are time-consuming and labor-intensive. Therefore, rapid AST approaches exist to address the delayed gap in time to actionable results. There are two main types of rapid AST technologies- phenotypic and genotypic approaches. In this review, we provide a summary of all commercially available rapid AST platforms for use in clinical microbiology laboratories. We describe the technologies utilized, performance characteristics, acceptable specimen types, types of resistance detected, turnaround times, limitations, and clinical outcomes driven by these rapid tests. We also discuss crucial factors to consider for the implementation of rapid AST technologies in a clinical laboratory and what the future of rapid AST holds.

Genome editing using type I-E CRISPR-Cas3 in mice and rat zygotes

The type I CRISPR system has recently emerged as a promising tool, especially for large-scale genomic modification, but its application to generate model animals by editing zygotes had not been established. In this study, we demonstrate genome editing in zygotes using the type I-E CRISPR-Cas3 system, which efficiently generates deletions of several thousand base pairs at targeted loci in mice with 40%-70% editing efficiency without off-target mutations. To overcome the difficulties associated with detecting the variable deletions, we used a newly long-read sequencing-based multiplex genotyping approach. Demonstrating remarkable versatility, our Cas3-based technique was successfully extended to rats as well as mice, even by zygote electroporation methods. Knockin for SNP exchange and genomic replacement with a donor plasmid were also achieved in mice. This pioneering work with the type I CRISPR zygote editing system offers increased flexibility and broader applications in genetic engineering across different species.

Comprehensive preimplantation genetic testing for balanced insertional translocation carriers

BackgroundBalanced insertional translocations (BITs) can increase the risk of infertility, recurrent miscarriages or neonatal birth defects due to chromosomal imbalances in gametes. However, studies on preimplantation genetic testing (PGT) for...

Genetic diversity of Plasmodium malariae in sub-Saharan Africa: a two-marker genotyping approach for molecular epidemiological studies.

Plasmodium malariae is the most common non-falciparum species in sub-Saharan Africa. Despite this, data on its genetic diversity is scarce. Therefore, we aimed to establish a P. malariae genotyping approach based on size polymorphic regions that can be easily applied in molecular epidemiological studies. Four potential genotyping markers, Pm02, Pm09, P. malariae thrombospondin-related anonymous protein (pmtrap), and P. malariae merozoite surface protein fragment 2 (pmmsp1 F2) were amplified via nested PCR and analysed using automated capillary gel electrophoresis. We observed the highest allelic diversity for pmtrap (MOI = 1.61) and pmmsp1 F2 (He = 0.81). Further applying the two markers pmtrap and pmmsp1 F2 on a different sample set of 21 P. malariae positive individuals followed up over one week, we saw a high consistency in their performance. The results show a large complexity and high dynamics of P. malariae infections in the asymptomatic Gabonese study population. We successfully implemented a new genotyping panel for P. malariae consisting of only two markers: pmtrap and pmmsp1 F2. It can be easily applied in other settings to investigate the genotype diversity of P. malariae populations, providing further important data on the molecular epidemiology of this parasite species.

Approach for Phased Sequence-Based Genotyping of the Critical Pharmacogene Dihydropyrimidine Dehydrogenase (DPYD).

Pre-treatment genotyping of four well-characterized toxicity risk-variants in the dihydropyrimidine dehydrogenase gene (DPYD) has been widely implemented in Europe to prevent serious adverse effects in cancer patients treated with fluoropyrimidines. Current genotyping practices are largely limited to selected commonly studied variants and are unable to determine phasing when more than one variant allele is detected. Recent evidence indicates that common DPYD variants modulate the functional impact of deleterious variants in a phase-dependent manner, where a cis- or a trans-configuration translates into different toxicity risks and dosing recommendations. DPYD is a large gene with 23 exons spanning nearly a mega-base of DNA, making it a challenging candidate for full-gene sequencing in the diagnostic setting. Herein, we present a time- and cost-efficient long-read sequencing approach for capturing the complete coding region of DPYD. We demonstrate that this method can reliably produce phased genotypes, overcoming a major limitation with current methods. This method was validated using 21 subjects, including two cancer patients, each of whom carried multiple DPYD variants. Genotype assignments showed complete concordance with conventional approaches. Furthermore, we demonstrate that the method is robust to technical challenges inherent in long-range sequencing of PCR products, including reference alignment bias and PCR chimerism.

Using a new analytic approach for genotyping and phenotyping chromosome 9p deletion syndrome.

Using a new analytic method ("unique non-overlapping region" (UNOR) analysis), we characterized the genotypes and phenotypes of a large cohort of individuals diagnosed with chromosome 9p deletion syndrome (9PMS) and defined critical genomic regions. We extracted phenotypic information from 48 individuals with 9PMS from medical records and used a guided interview with caregivers to clarify ambiguities. Using high-resolution whole-genome sequencing for breakpoint definition, we aligned deletions and drew virtual breakpoints to obtain UNORs associated with phenotypic characteristics. We next extracted genotype and phenotype data for 57 individuals identified from a systematic review of the 9PMS literature and analyzed these as above. Common phenotypic features included developmental delay/intellectual disability, dysmorphic features, hypotonia, genital defects in XY individuals, psychiatric diagnoses, chronic constipation, atopic disease, vision problems, autism spectrum disorder, gastroesophageal reflux disease, trigonocephaly, congenital heart disease, and neonatal hypoglycemia. Our approach confirmed previous literature reports of an association of FREM1 with trigonocephaly and suggested a possible modifier element for this phenotype. In conclusion, the UNOR approach delineated phenotypic characteristics for 9PMS and confirmed the critical role of FREM1 and a possible long-distance regulatory element in pathogenesis of trigonocephaly that will need to be replicated in future studies.

On leveraging self-supervised learning for accurate HCV genotyping

Hepatitis C virus (HCV) is a major global health concern, affecting millions of individuals worldwide. While existing literature predominantly focuses on disease classification using clinical data, there exists a critical research gap concerning HCV genotyping based on genomic sequences. Accurate HCV genotyping is essential for patient management and treatment decisions. While the neural models excel at capturing complex patterns, they still face challenges, such as data scarcity, that exist a lot in computational genomics. To overcome this challenges, this paper introduces an advanced deep learning approach for HCV genotyping based on the graphical representation of nucleotide sequences that outperforms classical approaches. Notably, it is effective for both partial and complete HCV genomes and addresses challenges associated with imbalanced datasets. In this work, ten HCV genotypes: 1a, 1b, 2a, 2b, 2c, 3a, 3b, 4, 5, and 6 were used in the analysis. This study utilizes Chaos Game Representation for 2D mapping of genomic sequences, employing self-supervised learning using convolutional autoencoder for deep feature extraction, resulting in an outstanding performance for HCV genotyping compared to various machine learning and deep learning models. This baseline provides a benchmark against which the performance of the proposed approach and other models can be evaluated. The experimental results showcase a remarkable classification accuracy of over 99%, outperforming traditional deep learning models. This performance demonstrates the capability of the proposed model to accurately identify HCV genotypes in both partial and complete sequences and in dealing with data scarcity for certain genotypes. The results of the proposed model are compared to NCBI genotyping tool.

Comprehensive Assessment of Bacterial Diseases in the Shrimp: Clinical, Phenotypic, Genotypic, and Histopathological Approaches

Comprehensive Assessment of Bacterial Diseases in the Shrimp: Clinical, Phenotypic, Genotypic, and Histopathological Approaches

Exploring immuno-genetic resistance in pigs to the food-borne zoonotic pathogen, Toxoplasma gondii: A serological study in Central Italy

Toxoplasmosis is a re-emerging food-borne zoonosis of warm-blooded animals caused by Toxoplasma gondii, an obligate intracellular parasite with a complex biological cycle. Until now, the genotypic approach for discovering putative resistance immuno-genetic markers has never been adopted for T. gondii infection. Thus, this study aimed, for the first time, to deepen these novel insights. In particular, pigs, reared in different farms in Central Italy, were phenotypically characterized by serological assays performed on diaphragm meat juice. Out of 179 tested animals, 98 resulted seropositive (54.74%), 57 seronegative (31.84%), and 24 doubtful (13.40%), underlining a possible re-emerging diffusion of this protozoan in the investigated areas. Contextually, an RT-PCR assay, followed by Sanger sequencing for IL-18, a pro-inflammatory cytokine with a key role upstream of the infection, was developed. At this stage, for this interleukin, significant polymorphic variations were not detected compared to the reference sequence, except in a seropositive animal. However, the starting outcomes of this novel and preliminary study will be investigated in depth with different approaches also on other target genes, with a crucial activity in the immunity pathway towards T. gondii, to unveil the possible presence of resistance genetic mechanisms and, finally, to make pork derived food safer.

Multimodal Learning for Mapping the Genotype-Phenotype Dynamics.

How complex phenotypes emerge from intricate gene expression patterns is a fundamental question in biology. Quantitative characterization of this relationship, however, is challenging due to the vast combinatorial possibilities and dynamic interplay between genotype and phenotype landscapes. Integrating high-content genotyping approaches such as single-cell RNA sequencing and advanced learning methods such as language models offers an opportunity for dissecting this complex relationship. Here, we present a computational integrated genetics framework designed to analyze and interpret the high-dimensional landscape of genotypes and their associated phenotypes simultaneously. We applied this approach to develop a multimodal foundation model to explore the genotype-phenotype relationship manifold for human transcriptomics at the cellular level. Analyzing this joint manifold showed a refined resolution of cellular heterogeneity, enhanced precision in phenotype annotating, and uncovered potential cross-tissue biomarkers that are undetectable through conventional gene expression analysis alone. Moreover, our results revealed that the gene networks are characterized by scale-free patterns and show context-dependent gene-gene interactions, both of which result in significant variations in the topology of the gene network, particularly evident during aging. Finally, utilizing contextualized embeddings, we investigated gene polyfunctionality which illustrates the multifaceted roles that genes play in different biological processes, and demonstrated that for VWF gene in endothelial cells. Overall, this study advances our understanding of the dynamic interplay between gene expression and phenotypic manifestation and demonstrates the potential of integrated genetics in uncovering new dimensions of cellular function and complexity.

High-throughput sequencing of Strongyloides stercoralis - a fatal disseminated infection in a dog.

The rhabditid nematode Strongyloides stercoralis is known worldwide as the causative agent of strongyloidiasis in humans. In addition to public health concerns, S. stercoralis also infects dogs, which represent a possible reservoir for potentially zoonotic transmissions. We describe the first confirmed case of fatal disseminated infection in a dog in the Czech Republic. The microscopic and histological results were supported by a complex genotyping approach. Using high-throughput sequencing of the hypervariable region (HVR-IV) of 18S rDNA and Sanger sequencing of the partial cytochrome c oxidase subunit 1 gene (cox1), the potentially zoonotic haplotype/lineage A of S. stercoralis was confirmed, while the solely canine haplotype/lineage B was not found. The development of the disease is mainly associated with immunodeficiency, and in this case, it was triggered by inappropriate treatment, in particular the use of corticosteroids.

An innovative single-cell approach for phenotyping and functional genotyping of CAR NK cells

BackgroundTo accelerate the translation of novel immunotherapeutic treatment approaches, the development of analytic methods to assess their efficacy at early in vitro stages is necessary. Using a droplet-based microfluidic platform,...

Investigation of Carbapenemase Production Among Carbapenem-Resistant Pseudomonas aeruginosa Isolates

Background: Pseudomonas aeruginosa is an opportunist organism that causes potentially life threatening nosocomial infections, particularly in immunocompromised patients. Carbapenems are regarded to be the last line of treatment against severe infections caused by multi drug resistant P. aeruginosa isolates. isolates. Production of the carbapenemase enzyme is the primary mechanism of carbapenem resistance and has become a serious health concern worldwide as these enzymes are highly transferable and limit therapeutic alternatives. Rapid detection of carbapenemase production is important for prompt planning the treatment of car-bapenemase-producing isolates and preventing the spread of these strains. This study aimed to investigate carbapenemase produc-tion in carbapenem resistant Pseudomonas aeruginosa isolates by the Carbapenem inactivation method. Materials and Methods: In this retrospective study a total of 172 Pseudomonas aeruginosa isolates were obtained from different samples sent from various clinics to Tokat Gaziosmanpaşa University Research and Application Hospital Microbiology Laboratory between 2016-2019 and were evaluated. Of the 172 isolates, 51 (29.7%) were found to be carbapenem- resistant and included in this investigation. Identification and antibiotic susceptibility tests of the isolates were performed with the Vitek 2 (Biomerieux, France) automated system. Carbapenem sensitivities were also determined by the disc diffusion method. Carbapenemase production in isolates was investigated by the Carbapenem inactivation method. Results: These samples were sent from clinical units, such as neurology (n =10), general surgery (n =8), internal medicine (n =7), and pediatric (n =6). The isolates were identified from wounds (n = 17), sputum (n = 15), blood (n = 11), urine (n = 5), and cerebrospinal fluid (n = 3) samples. Of all the carbapenem –resistant samples 32 (62.8%) were obtained from male, and 19 (37.3%) from female patients. Of the 51 carbapenem resistant isolates, 38 (74.5%) were found to be resistant to both imipenem and meropenem. Eight (15.7%) isolates were found to be resistant to imipenem only, and five (9.8%) isolates were resistant to meropenem. Carbapenemase production was detected in 31 (60.8%) isolates by using using the Carbapenem inactivation method. The antibiotic resistance rates of the carbapenem-resistant isolates were as follows: piperacillin-tazobactam 65%, amikacin 6.8%, gentamicin 15.2%, ceftazidime 34.6%, cefepime 38.3%, ciprofloxacin 26.7%, levofloxacin 24.2%. Conclusions: Rapid identification of carbapenemase enzymes among carbapenem resistant Pseudomonas aeruginosa isolates using phenotypic and genotypic approaches is important to control the transmission of infection caused by carbapenem-resistant isolates and to control the morbidity and mortality associated with them. In this study, the carbapenem inactivation test was seen as a method that can be preferred in the laboratory in terms of its easy and fast application in the detection of carbapenemase production. Key Words: Pseudomonas aeruginosa, Carbapenem inactivation method, Antimicrobial resistance