RESUMEN
The non-tuberculosis mycobacteria Mycobacterium avium subsp. hominissuis (MAH) are able to cause human mycobacteriosis. In this work, the results of the first comprehensive study of the genome polymorphism of the clinical strains of MAH were reported using the typing scheme by 13 loci MATR-VNTR (TR292, TRX3, TR25, TR47, MATR-1, MATR-4, MATR-5, MATR-6, MATR-8, MATR-11, MATR-14, MATR-15, MATR-16) containing tandem nucleotide sites and IS1245-RFLP-typing sites. A total of 90 MAH strains isolated from patients with lung mycobacteriosis without epidemiological connection (including HIV infected) were tested in 2008-2011. The inhomogeneity of the MAH strains by 36 profiles of 13 loci MATR-VNTR was observed. The majority of the strains (68.8%) were included in the 8 MATR-VNTR clusters; most large cluster contained 37 strains with 13-bitnumerical profile 2222223145443'. The nucleotide sequence of the MATR-16 (3') locus contains the long deletion (GenBank accession no. KF479191). The MAH strains of the MATR-VNTR clusters were found to be inhomogeneous by the IS1245 marker. The MATR-VNTR-typing method by 13 loci is recommended for preliminary differentiation of domestic MAH strains with further analysis of the MATR-VNTR clusters using the IS1245-RFLP-typing method.
Asunto(s)
Genoma Bacteriano , Infecciones por Mycobacterium no Tuberculosas/genética , Mycobacterium avium/genética , Secuencias Repetidas en Tándem/genética , Secuencia de Bases , VIH/genética , VIH/patogenicidad , Humanos , Mycobacterium avium/patogenicidad , Filogenia , Polimorfismo de Longitud del Fragmento de Restricción/genéticaRESUMEN
The DNA of virus of human papilloma of high carcinogenic risk was detected in 116 cervical samples. At that, the morphological symptoms of background processes are detected in 19 samples, CIN 1 in 9, CIN 2 in 23, CIN 3 in 54 (and out of them carcinoma in situ in 13), epidermoid cancer (squamous cell carcinoma) in 11 cases. The viral load of human papilloma of high carcinogenic risk in all samples of DNA exceeded threshold of clinical value (3 lg copies of DNA of human papilloma/105 cells). The genetic typing of human papilloma of high carcinogenic risk revealed the dominance of human papilloma of type 16 in 49.7%, type 33 in 15.3%, type 31 in 12.3% and type 45 in 5.5%. In women with background processes in cervix of the uterus DNA of human papilloma type 16 was detected more often in episome form. In case of dysplastic alterations of epithelium and cervical cancer DNA of human papilloma type 16 is detected in mixt form with different degree of integration into cell genome.
Asunto(s)
Carcinoma de Células Escamosas/virología , ADN Viral/metabolismo , Epitelio/virología , Papillomavirus Humano 16/metabolismo , Infecciones por Papillomavirus/virología , Reacción en Cadena de la Polimerasa/métodos , Neoplasias del Cuello Uterino/virología , Adulto , Anciano , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patología , Cuello del Útero , ADN Viral/genética , Epitelio/metabolismo , Epitelio/patología , Femenino , Papillomavirus Humano 16/genética , Humanos , Persona de Mediana Edad , Infecciones por Papillomavirus/genética , Infecciones por Papillomavirus/metabolismo , Infecciones por Papillomavirus/patología , Neoplasias del Cuello Uterino/genética , Neoplasias del Cuello Uterino/metabolismo , Neoplasias del Cuello Uterino/patologíaRESUMEN
Sixty 60 human papillomavirus (HPV) type 16-positive cervical epithelium biopsy specimens were studied by cytological, histological, immunomorphological, and PCR assays. No correlation was found between proliferation markers and the degree of integration of HPV 16 DNA integration into the cell genome. The number of cells with Ki-67 expression, mitoses, and their alteration was related to the grade of epithelial lesion. Expression of p53 was detected only in the episomal form of HPV DNA and its low integration.
Asunto(s)
Cuello del Útero , ADN Viral/metabolismo , Genoma Humano , Papillomavirus Humano 16/metabolismo , Infecciones por Papillomavirus , Integración Viral , Adulto , Anciano , Cuello del Útero/metabolismo , Cuello del Útero/patología , Cuello del Útero/virología , Epitelio/metabolismo , Epitelio/patología , Epitelio/virología , Femenino , Regulación de la Expresión Génica , Humanos , Antígeno Ki-67/biosíntesis , Persona de Mediana Edad , Infecciones por Papillomavirus/metabolismo , Infecciones por Papillomavirus/patología , Infecciones por Papillomavirus/virología , Proteína p53 Supresora de Tumor/biosíntesisRESUMEN
AIM: Molecular-genetic characteristic of M. tuberculosis strains isolated from operation material of patients with tuberculous spondylitis. MATERIALS AND METHODS: 107 strains of M. tuberculosis isolated in 2007 - 2011 from patients with spine tuberculosis were studied by methods of spoligotyping and MIRU-VNTR by 12 and 24 loci. RESULTS: Strains of genetic family Beijing dominated (n = 80), 78% of those had multiple drug resistance (MDR). Strains of genetic families T, H3 (Ural), LAM, Manu, H4 and S were also detected. Differentiating of 80 strains of Beijing genotype by MIRU-VNTR method by 24 loci revealed 24 variants (HGI = 0.83) including 7 clusters, the largest of those (100-32) included 23 strains (87% MDR). CONCLUSION: The leading role of Beijing genotype M. tuberculosis strains in development of tuberculous spondylitis with multiple drug resistance of the causative agent is shown.
Asunto(s)
Sitios Genéticos , Variación Genética , Mycobacterium tuberculosis/genética , Espondilitis/genética , Tuberculosis Resistente a Múltiples Medicamentos/genética , Tuberculosis de la Columna Vertebral/genética , Femenino , Humanos , Masculino , Mycobacterium tuberculosis/aislamiento & purificación , Espondilitis/microbiología , Tuberculosis de la Columna Vertebral/microbiologíaRESUMEN
Mycobacterium avium are typical environmental, non-tuberculosis microorganisms that occasionally cause mycotuberculosis, an infectious disease in wild and domestic animals, birds, and humans. Here, we report the results of the first study on the genetic diversity of the Russian population of M. avium. A total of 85 M. avium subsp. hominissuis (MAH) clinical strains were isolated from patients (including 30 HIV-positive individuals) with mycobacteriosis in St. Petersburg, 2008-2011. The biochemical identification of the microorganisms was carried out using the PCR detection of the mobile elements IS901 and IS900, as well as of the polymorphism of restriction fragments of the hsp65 gene. The genetic diversity of the isolates was evaluated by VNTR typing based on eight variable-number tandem repeats (VNTRs) (292, X3, 25, 47, 3, 7, 10, and 32 [Thibault et al., 2007]). The MAH population studied was characterized by 15 VNTR types, including nine unique patterns and six clusters of isolates with identical eight-digit profiles. The largest clusters (22221128 and 24221128) included 45 (59.2%) and 15 (19.7%) isolates, respectively; the others contained from 2-7 strains. The strains of the cluster 2533112'8 possessed a truncated TR10 locus (allele 2'). Taking into account the absence of the epidemiological links between the patients and the fact that the infection was delivered from the environment, the high rate of clustering of MAH isolates can be explained by the low discriminatory power of the eight-locus VNTR-typing scheme (HGDI 0-0.61).
Asunto(s)
Variación Genética , Repeticiones de Minisatélite , Mycobacterium avium/genética , Proteínas Bacterianas/genética , Chaperonina 60/genética , Genotipo , Humanos , Mycobacterium avium/aislamiento & purificación , Polimorfismo Genético , Polimorfismo de Longitud del Fragmento de Restricción , Federación de RusiaRESUMEN
AIM: Characteristics of drug resistance (DR) and population structure of Mycobacterium tuberculosis in Pskov region. MATERIALS AND METHODS: In 90 strains of M. tuberculosis drug resistance was studied by culture method and by using "TB-BIOCHIP"; genotyping was determined by spoligotyping method. RESULTS: 55 (61.1%) of 90 M.tuberculosis strains had drug resistance, with 40 (44.4%) being multi-resistant. M. tuberculosis population was presented by SIT1 spoligotype strains of genetic families Beijing--44.4%, LAM--21.1%, T--14.4%, Haarlem--11.1% and Ural--5.6%, according to SpolDB4. Among M. tuberculosis strains circulating in Pskov region the most widespread (44.4%) was SIT1 spoligotype (p < 0.0001). DR and multi-resistant DR (MDR) in Beijing strains occurred more frequently than in "non-Beijing" strains (p < 0.001 and p = 0.03 respectively) and were determined by rpoB mutations Ser531-->Ley and katG Ser315-->Thr. All the SIT252 spoligotype strains were multi-resistant, and their resistance to rifampicin was determined by rpoB Asp516-->Ser substitution, to isoniazid --katG Ser315-->Thr and inhA_T15 substitutions. CONCLUSION: The data obtained gives evidence on tuberculosis epidemiological unfavorability and wide circulation of MDR M. tuberculosis strains in Pskov region.
Asunto(s)
Antituberculosos/administración & dosificación , Farmacorresistencia Bacteriana Múltiple/genética , Genes Bacterianos , Mycobacterium tuberculosis/genética , Tuberculosis Resistente a Múltiples Medicamentos/microbiología , Adulto , Anciano , Antituberculosos/uso terapéutico , Técnicas de Tipificación Bacteriana , Femenino , Genotipo , Humanos , Isoniazida/administración & dosificación , Isoniazida/uso terapéutico , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Mutación , Mycobacterium tuberculosis/clasificación , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/aislamiento & purificación , Rifampin/administración & dosificación , Rifampin/uso terapéutico , Federación de Rusia/epidemiología , Tuberculosis Resistente a Múltiples Medicamentos/tratamiento farmacológico , Tuberculosis Resistente a Múltiples Medicamentos/epidemiologíaRESUMEN
AIM: Study of specter of low-manifest infections (LMI) with central nervous system (CNS) damage and their role in patients in prolonged unconscious state (PUS) of noninflammatory etiology. MATERIALS AND METHODS: 32 patients (23 male, 9 female; age 14-58) in PUS of various etiology were examined. The main group (18 patients) received therapy against all infectious diseases including LMI; control group (14 patients)--only against common and nosocomial microflora. Patients were immunologically, infectologically and neurologically examined in dynamic. The data obtained were treated by using STATISTICA for Windows (version 5.5). RESULTS: Significant differences in immune and infectologic status depending on the nature of primary CNS damage were not detected. Immunodeficiency was detected in all patients; 94% of patients had increased non-specific IgM and IgE. Among LMI agents Chlamydia spp. were predominant. Cultural and/or PCR methods detected this microorganism during the primary examination in cerebrospinal fluid samples in 56% patients and in blood samples in 56%; during the second diagnostics or autopsy--only in 13 and 25%, respectively. Detection of Bacteroides fragilis, Human Herpes Virus (HHV-6), Virus Epstein Barr (VEB), Cytomegalovirus (CMV) in cerebrospinal fluid, blood and on mucous membranes of nasopharynx and conjunctiva was grouped more frequently with the presence of Chlamydia spp. in the CNS (p < 0.05) than with other LMI agents. Sanation of CNS from LMI was significantly accompanied by regeneration of communicative activity in comparison with the control group. CONCLUSION: In patients with PUS high frequency of CNS infection by various LMI agents and primarily Chlamydia spp. should be considered. Sanation from LMI can become a "window" for effective neuro-regenerative treatment.
Asunto(s)
Infecciones Bacterianas/diagnóstico , Infecciones del Sistema Nervioso Central/diagnóstico , Chlamydia/aislamiento & purificación , Coinfección , Inconsciencia/diagnóstico , Virosis/diagnóstico , Adolescente , Adulto , Antibacterianos/administración & dosificación , Antivirales/administración & dosificación , Infecciones Bacterianas/líquido cefalorraquídeo , Infecciones Bacterianas/tratamiento farmacológico , Infecciones Bacterianas/microbiología , Infecciones Bacterianas/virología , Bacteroides fragilis/genética , Bacteroides fragilis/aislamiento & purificación , Sistema Nervioso Central/microbiología , Sistema Nervioso Central/patología , Sistema Nervioso Central/virología , Infecciones del Sistema Nervioso Central/líquido cefalorraquídeo , Infecciones del Sistema Nervioso Central/tratamiento farmacológico , Infecciones del Sistema Nervioso Central/microbiología , Infecciones del Sistema Nervioso Central/virología , Chlamydia/genética , Citomegalovirus/genética , Citomegalovirus/aislamiento & purificación , ADN Bacteriano/análisis , ADN Viral/análisis , Femenino , Herpesvirus Humano 4/genética , Herpesvirus Humano 4/aislamiento & purificación , Herpesvirus Humano 6/genética , Herpesvirus Humano 6/aislamiento & purificación , Humanos , Masculino , Persona de Mediana Edad , Tipificación Molecular , Reacción en Cadena de la Polimerasa , Inconsciencia/líquido cefalorraquídeo , Inconsciencia/tratamiento farmacológico , Inconsciencia/microbiología , Inconsciencia/virología , Virosis/líquido cefalorraquídeo , Virosis/tratamiento farmacológico , Virosis/microbiología , Virosis/virologíaRESUMEN
AIM: Improvement of etiologic diagnostics of disseminated lung tuberculosis (DLT) and determination of Mycobacterium tuberculosis (MBT) drug susceptibility on the basis of molecular genetic methods. MATERIALS AND METHODS: Samples from respiratory tract of patients with DLT were studied using real time polymerase chain reaction and the "TB-BIOCHIP" assay developed by Institute of Molecular Biology. Methods of spoligotyping and reverse hybridization were used for identification, genotyping and express-detection of drug resistance of MBT to rifampicin in sputum samples stained for bacterioscopy. RESULTS: In 76 (41.5%) of 183 patients with radiological signs of DLT, DNA of tuberculosis complex mycobacteria was detected in respiratory tract samples (specificity 87.7%); mutations in genes rpoB, katG, inhA as well as region ahpC-oxyR associated with resistance to rifampicin and isoniazide were revealed in 67% and 79.5% of patients with DLT respectively. In 48.8% of sputum samples, DNA of MBT of epidemically significant genotype Beijing associated with multidrug resistance of MBT in Russia was identified. CONCLUSION: Molecular genetic methods allow to use both fresh and archived respiratory tract specimens for rapid verification of DLT diagnosis during oligobacillar forms of tuberculosis as well as timely prescribe and correct the treatment regimen of the patient according to individual drug susceptibility spectrum of the agent.
Asunto(s)
Mycobacterium tuberculosis/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Tuberculosis Pulmonar/diagnóstico , Antibióticos Antituberculosos/farmacología , Proteínas Bacterianas/genética , Catalasa/genética , ARN Polimerasas Dirigidas por ADN , Diagnóstico Diferencial , Farmacorresistencia Bacteriana , Genes Bacterianos/genética , Humanos , Mutación/efectos de los fármacos , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/genética , Oxidorreductasas/genética , Rifampin/farmacología , Federación de Rusia , Esputo/microbiologíaRESUMEN
The natural wild rhizosphere strain P. fluorescens 2137 was marked with the beta-glucuronidase gene gusA. The introduction of this gene influenced the viability of the wild strain, as well as its certain physiological parameters, such as cultural characteristics, biochemical properties, and antagonistic activity against the phytopathogenic fungi Fusarium culmorum, F. oxysporum, F. graminearum, and Verticillum nigrescens. The gusA-marked derivative strains that deviate the least from the wild strain in biological properties can be used to monitor populations of P. fluorescens 2137 cells in the plant rhizosphere.
Asunto(s)
Glucuronidasa/fisiología , Pseudomonas fluorescens/fisiología , Antibiosis , Medios de Cultivo , Hongos/inmunología , Glucuronidasa/genética , Plantas/microbiología , Pseudomonas fluorescens/enzimología , Pseudomonas fluorescens/genética , Microbiología del Suelo , Transformación BacterianaRESUMEN
The paper presents the microbiological and molecular genetic characteristics of Mycobacterium tuberculosis (MBT) isolated from multiple lesion foci in 26 patients with multiorgan and generalized tuberculosis. Cultures of MBT of the family Beijing were isolated from the pathological specimens taken from 17 (65.4%) patients; those with individual genotypes were in 9 (34.6%) patients. The study indicated that cultured MBT from different types of pathological material from 25 of the 26 patients had identical spoligotypes irrespective of their clinical diagnosis. At the same time minor profile IS6110 changes might occur in the Mycobacteria of a patient during persistence. It may be concluded that multiple foci of tuberculous affliction in the patients are of unique origin and have the mechanism of development, associated with endogenous reactivation of infection.