[Comparison of efficacy of the rapid mycobacteria growth indicator tube (MGIT) culture method (manual system) and conventional culture method in mycobacterial detection]. / Posouzení ucinnosti metabolické kultivacní rychlometody MGIT (manuální systém) pro záchyt mykobakterií ve srovnání s klasickou kultivacní metodou.

Diagnosis of tuberculosis is based on clinical symptoms, lung X-ray, skin tuberculin test and primarily on the detection of the causative agent Mycobacterium tuberculosis by direct microscopy of biological specimens and culture on solid egg media and in liquid media (Ogawa, Löwenstein-Jensen, Sula). Slow growth of most mycobacterial species is a limiting factor in both the confirmation of etiology and subsequent drug susceptibility tests and species identification that are of crucial relevance to early institution of treatment, selection of treatment regimen and implementation of antiepidemic measures. One of the methods proposed for more rapid detection of mycobacteria and suitable for use in routine diagnostic laboratories is the BD BBL MGIT culture system (Becton Dickinson, 1 Becton Drive, Franklin Lakes, NJ 07417, USA) based on fluorescence detection of the initial phase of mycobacterial multiplication at which macrocolony growth is still not visible. The fluorescent compound Tris, 4,7-diphenyl-1,10-phenalthroline ruthenium chloride pentahydrate is embedded in silicone on the bottom of tubes with liquid culture medium in which growing, actively respiring mycobacteria consume the oxygen and allow the fluorescence to be detected and visualized using a UV transluminator.

[Nontuberculous mycobacteria and incidence of mycobacterioses in Prague in 1999-2004]. / Netuberkulózní mykobakteria a mykobakteriózy v Praze v letech 1999-2004.

OBJECTIVE: To analyze the incidence of nontuberculous mycobacteria detected in Prague patients in 1999-2004 as either single findings suggestive of clinical insignificance or repetitive findings suggestive of mycobacteriosis when reported together with the corresponding organ damage and symptomatology. MATERIAL AND METHODS: Nontuberculous mycobacteria were isolated and identified according to the Czech recommended methods for mycobacterial diagnosis in two Prague laboratories. The determined incidence rates of Mycobacterium (M.) kansasii, M. avium complex (MAC) and M. xenopi were compared with the respective nationwide rates and those of other nontuberculous bacteria, M. tuberculosis and M. bovis. The data on cases of mycobacterioses reported within the Czech Registry of Tuberculosis were provided by the Institute of Health Information and Statistics of the Czech Republic. RESULTS: In 1999-2004, the annual incidence rates of nontuberculous mycobacteria detected in Prague patients ranged between 169-139 and accounted for 13-25% of the totals of isolated mycobacteria including M. tuberculosis and M. bovis. Over this period, M. kansasii, MAC and M. xenopi were detected in 45, 76 and 43 patients, respectively. The single to repetitive detection ratio was the highest for M. kansasii (1:1.6), followed by MAC (1:1.8) and M. xenopi (1:2.3). Most male excretors were from higher age categories (median of 58-73 years) while the median age of female excretors ranged between 41 and 75 years. As many as 47 cases of mycobacterioses caused by the following agents: M. kansasii (20 cases), MAC (17 cases), M. xenopi (6 cases) and other nontuberculous mycobacteria (4 cases), were reported to the National Registry of Tuberculosis over the study period. CONCLUSIONS: Detection and identification of nontuberculous mycobacteria have become part of diagnostic routine of mycobacteriological laboratories. Compared to conventional tuberculous mycobacteria, detection of nontuberculous mycobacteria often requires the use of different and more cumbersome procedures such as incubation at preferential temperatures, longer incubation for detection of growth in primary cultures, species specific culture media, etc.. More skills and experience are needed for the use of automated detection systems and molecular biological techniques for species identification and interpretation of results. Regular consultations with clinical and outpatient physicians are crucial for the assessment of pathogenetic potential of nontuberculous mycobacteria.

Molecular epidemiology of tuberculosis in Prague and South Moravia, Czech Republic: genetic analysis of Mycobacterium tuberculosis isolates by IS6110-RFLP fingerprinting and spoligotyping.

OBJECTIVES: To genetically characterize and compare Mycobacterium tuberculosis isolates among culture-confirmed TB cases in two regions in the Czech Republic in 1998. METHODS: Consecutive M. tuberculosis isolates from 111 TB patients in Prague and 120 patients in the South Moravia region were genotyped using the standardized IS6110 Southern blot hybridization method and by spoligotyping. RESULTS: Eighty of the Prague patients (72.1%) had isolates with unique RFLP patterns, while 31 (27.9%) had isolates which belonged to 10 clusters. Seventy-eight (64.7%) of the South Moravia strains displayed unique RFLP pattern and 42 (35.3%) were assigned into 15 clusters. The spoligotype profiles previously identified in the U.S. were found in 69 (33%) samples and newly identified Czech spoligotypes in 24 (11.4%) of the total number of examined strains. CONCLUSIONS: The present population-based molecular epidemiological study performed in two regions of the Czech Republic in 1998 demonstrated the distribution of individual genotypes as well as clustered strains of M. tuberculosis isolated from TB patients, and confirmed the similarity between the Czech strain collection and the European Community TB Database, that includes countries with low TB rate. The sporadic import of TB cases from foreign countries and recent transmission events probably do not play significant roles in the epidemiological situation in the Czech Republic.

[Molecular epidemiology of tuberculosis among prisoners]. / Molekulární epidemiologie tuberkulózy veznu.

The subject of the investigation is a group of 27 strains of Mycobacterium tuberculosis isolated in 2000 from prisoners with the diagnosis of tuberculosis, 19 Czechs and 8 foreigners (mean age 41 and 35 years resp.). The molecular-epidemiological examination of these strains was made using the RFLP fingerprint technique (Restriction Fragment Length Polymorphism) with evidence of the insertive sequence IS6110 and the technique of spoligotyping, based on detection of hybridization of spacer oligonucleotides. DNA fingerprinting revealed a high polymorphism in the number and molecular weight of sequence IS6110 which is common in Czech and other European strains of M. tuberculosis. All strains with the exception of two had mutually different fingerprint profiles. In the two with identical fingerprints probably a duplicit examination of the same material was involved which occurred by mistake during transport or in the laboratory. The fingerprint method thus did not prove interhuman transmission of tuberculosis between the examined prisoners. The technique of spoligotyping revealed the finding of genotype Beijing M. tuberculosis in two sick prisoners, one Algerian and one Albanese, and in one Czech prisoner. This genotype found in a high percentage of patients in southeastern Asia and in migrants from this area was detected for the first time in the Czech Republic. The findings are evidence of a satisfactory standard of the programme of tuberculosis control in the Czech prison system and at the same time draw attention to the potential possibility of the spread of tuberculosis from migrants coming from areas with a high prevalence.

[Spoligotypes of Mycobacterium tuberculosis in the Czech population (methods and pilot study)]. / Spoligotypy ceských populací Mycobacteria tuberculosis (metodické principy a pilotní studie).

In the introduction the authors describe molecular genetic principles of spacer oligonucleotide typing of the M. tuberculosis complex and mention in detail the methodical procedure of implementation of this examination technique incl. computer analysis of results. In the pilot part of the study the authors describe examination of 71 strains of M. tuberculosis isolated from patients living in the capital of Prague and 37 strains from patients in the South Moravian region of the Czech Republic. In the group of Prague strains a total of 7 clusters was identified formed by 2-14 identical profiles of spoligotypes and 37 individually different profiles. In the South Moravian strains there were 6 clusters with 2-14 identical spoligotypes and 10 non-clustered profiles. In neither of the groups there were spoligotypes typical for the genetic family Beijing or Africa. The study is a preliminary molecular epidemiological analysis where DNA fingerprint RFLP profiles are compared with spoligotypes of strains of M. tuberculosis of patients from the capital of Prague and the South Moravian region of the Czech Republic.

[Human transmission of tuberculosis confirmed by DNA fingerprinting]. / Interhumánní prenos tuberkulózy potvrzený fingerprintingem DNA.

By means of fingerprinting based on detection of the insertive sequence IS6110 v DNA of M. tuberculosis predigested by the restrictive enzyme PVUII the authors proved epidemiological associations in three patients with bacillary tuberculosis. The primary source of infection was a 44-year-old man, alcoholic and homeless suffering from tuberculosis diagnosed during an episode of ethylic ebriety, discharged prematurely from hospital because of lack of discipline and drunkenness who died after 8 months with a finding of caseous pneumonia. The first contact person was a 53-year-old women suffering from diabetes and hypertension, an auxiliary worker in the laboratory where she probably was infected during manipulation of the patients infected sputum. The other patients was a 49-year-old man, alcoholic, unemployed where a symptomatic tuberculous finding was detected with general weakness, elevated temperatures and expectoration, who most probably was infected by a patient during accidental contact in his domicile. The profiles of RFLP strains of M. tuberculosis of all three patients had an identical appearance of the DF-1 clone formed by seven copies of IS6110. This profile was not found in any of the total of 98 strains of M. tuberculosis examined within the framework of the molecular epidemiological prevalence study of patients with bacillary tuberculosis notified in Prague in 1999.

[Stomatologic care of patients with the Melkersson-Rosenthal syndrome]. / Stomatologická péce o pacienty s Melkersonovým-Rosenthalovým syndromem.

The authors describe the problems of diagnosis, treatment, prevention and dispensarization of patients suffering from Melkerson-Rosenthal's syndrome in a dental department. They describe in detail the manifestations of the syndrome in the orofacial area. Comprehensive dental care is based on studies of the pertinent literature and the authors' own experience.

[Complex stomatologic care in patients with anorexia nervosa]. / Komplexní stomatologická péce o nemocné s mentální anorexií.

The authors describe a group of 25 patients with anorexia nervosa treated at the Psychiatric Clinic, First Medical Faculty, Charles University. They compare their experience with dental treatment of these patients with data in the literature. Extensive dental damage in these patients calls for comprehensive dental care which is not only time consuming but frequently also pretentious from the economic aspect. The authors recommend therefore dispensary care and emphasize the necessity of collaboration between the psychiatrist and stomatologist.

Trehalase activity in diabetes mellitus and in cirrhosis of the liver.

Trehalase (an enzyme decomposing the disaccharide trehalose) activity was studied in 29 healthy subjects, 25 patients with cirrhosis and 112 diabetics. Mean trehalase activity was 176 +/- 11 units in the control group, 647 +/- 421 units in the patients with cirrhosis and 467 +/- 239 units in diabetics. The differences between the control group on the one hand and the groups with cirrhosis and diabetes on the other were statistically significant. The results show that the organism, under pathological conditions, makes far greater use of its enzymatic apparatus to assure its basic requirements, but the scatter of the values is so great that the determination of trehalase has no discriminative value in individual cases.