Cytogenetic and molecular genetic methods for chromosomal translocations detection with reference to the KMT2A/MLL gene

Acute leukemia (ALS) are often associated with chromosomal translocations, including KMT2A / MLL genes rearrangements. The identification or confirmation of these translocations is carried out by a number of genetic and molecular methods, some of which are regularly used in clinical practice, while others are mainly used for research purposes. In the clinic, these methods clarify diagnoses and monitor the course of illness and therapy. On the other hand, the identification of new translocations and the confirmation of known translocations is of paramount importance in the study of the disease mechanisms and an additional molecular classification.

There are multiple methods for detecting rearrangements that differ by their operating principle, the type of problem solved and the cost income statement. This opinion is intended to help researchers and clinicians studying al and associated chromosomal translocations to navigate this variety of methods. All the methods considered in the examination are grouped by their principle of action and include curyotyping, in situ hybridization of fluorescence (fish) with integer chromosome probes or individual loca, PCR and reverse transcription methods, as well as high speed sequencing.

Another characteristic of the methods described is the type of problem solved. This may be the discovery of new rearrangements, the determination of unknown partner genes participating in the rearrangement or confirmation of the proposed rearrangement between the two genes. We consider that the specificities of the application, the basic principle of each method, as well as its advantages and disadvantages. To illustrate the application, examples of study of KMT2A / MLL gene rearrangements, one of the gene often rearranged in Al, are mentioned.

RNF213 Mutation of the gene in the circulating tumor DNA detected by a new generation targeted sequencing in the first-step lung cancer discrimination of pulmonary nodules

Background: Distinguishing early lung cancer of benign disease in pulmonary nodules, particularly opacity lesions (GGO), we evaluate NTTNA gene mutations in peripheral blood using new sequencing. Generation (NGS) targeted.

Methods: Patients with simple pulmonary nodules without mediastinous lymph nodes and symptoms that are difficult to diagnose by the CT chest and the lung cancer biomarker measurement in several medical centers have been entered in the study. All patients accepted mini-invasive surgery but refused preoperative biopsy. Geneous mutations in preoperative blood samples were detected by targeted Ngs. Mutations with significant differences between pulmonary tumors and benign lesions, as grouped by postoperative pathology, have been screened. Protein expression was determined by immunohistochemistry. The very expressed genes have been selected as biomarkers to verify mutations of peripheral blood.

Results: In the training game, the RNF213, KMT2D, CSMD3 and LRP1B genes were mutated more frequently in early lung cancer (27 cases) than in benign nodules (15 cases) (p <0.05). A high expression of the RNF213 gene in lung cancers and low expression in benign diseases were observed by immunohistochemistry. The RNF213 gene was mutated in 25% of the lung cancer samples in the validation assembly of 28 samples and showed a high specificity (100%). In patients with GGONS, RNF213 was mutated more frequently in early lung cancer compared to benign diseases (p <0.05).

 Cytogenetic and molecular genetic methods for chromosomal translocations detection with reference to the KMT2A/MLL gene
Cytogenetic and molecular genetic methods for chromosomal translocations detection with reference to the KMT2A/MLL gene

 

Detection of antibiotic resistance genes among multiple pseudomonas resistant to medicines, isolated from clinical sources in health facilities selected in Kwara State

Background: Pseudomonas Aeruginosa (P. Aeruginosa) is a common nosocomial pathogen that causes serious illnesses in many clinical and community contexts. P. aeruginosa strains are associated with increased morbidity, mortality and health care. The rapid emergence of antimicrobial resistance between these strains is a public health crisis. In addition, there is a shortage of data on the characterization of P. aeruginosa isolates from human clinical samples in the state of Kwara.

Objectives: The objectives are to study the occurrence of the metallo-β-lactamase enzyme, multiple antibiotics P. aeruginosa, among the clinical samples and the detection of antibiotic resistance genes.

Methods: Two hundred and thirty-five samples comprising 145 men and 90 females Human clinical specimens were collected from five selected health facilities within Kwara State, Nigeria. The samples were grown immediately using standard microbiological procedures. Several drug resistance models from microorganisms to different antibiotics have been determined using the Bauer Kirby disk broadcasting technique. The production of metallo β-lactamase has been determined using the test tape E and DNA samples of multi-strains resistant to P. aeruuginosa have been extracted and subjected to a polymerase chain reaction (PCR) for the Determination of resistant genes. Data have been submitted to descriptive statistics using statistical software on the Social Science Package (SPSS).

Results: A total of 145 isolates were identified for P. aeruginosa .Chotorty clinical samples were positive for the production of metallo-β-lactamase; 11 (8%) men and 19 (13%) women. Absolute resistance to ceftazidime (100%), gentamicin (100%), ceftiaxone (100%) were observed while low ciprofloxacin resistance (12.4%), piperacillin (6.9%) and L IMIPENEM (6.9%). All isolates were sensitive to colistine. The prevalence of various Blavim coding genes, BLACX-M and Blatemem were 34.4%, 46.7%, 16.7% and 37.7% respectively.

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  • 10 plates of 96 wells
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Conclusion: This study showed that there was a high occurrence of metallo-β-lactamase enzymes producing and antibiotic pertainers of P. aeruginosa in clinical specimens of the studied area. The necessary measures must therefore be implemented to stop the problems of this antibiotic resistance.

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