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M Kubín

Publications and source records attributed to M Kubín.

At least 19 recordsLinked to original sources
PubMed2004

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.

Adult↗
PubMed2003

[Molecular epidemiology of tuberculosis among prisoners].

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.

Adult↗
PubMed2002

[Spoligotypes of Mycobacterium tuberculosis in the Czech population (methods and pilot study)].

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.

Bacterial Typing Techniques↗
PubMed2001

[DRE-PCR (Double Repetitive Element-Polymerase Chain Reaction)--a new molecular-epidemiologic method in the detection of tuberculosis].

In a pilot study Double Repetitive Element-Polymerase Chain Reaction 20 clinical isolates of Mycobacterium tuberculosis from Guatemala and 49 strains from Prague were typed. This technique is based on direct evidence of repetitive elements IS6110 or PGRS and does not require DNA purification, digestion by endonuclease nor Southern blot hybridization. Preliminary examination of Guatemalian strains revealed a striking identity or similarity of DRE-PCR profiles while the Prague strains were characterized by conspicuous polymorphism. The Prague strains were examined in a total number of 13 series of electrophoreograms and subsequently subjected to automated analysis with GelCompar software. The DRE-PCR method is suitable for screening of a major number of clinical isolates of M. tuberculosis in laboratories equipped with a minimum of technical facilities for the PCR method, furthermore it requires facilities for synthesis of the necessary primers and at least basic knowledge of molecular biology.

Bacterial Typing Techniques↗
PubMed2001

[Human transmission of tuberculosis confirmed by DNA fingerprinting].

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.

Adult↗
PubMed2000

The first occurrence of a multi-drug resistant tuberculosis epidemic in the Czech Republic caused by genetically closely related Mycobacterium tuberculosis strains.

DNA fingerprinting based on the detection of the insertion sequence IS6110 in Pvull restriction fragments was applied to M. tuberculosis isolates originating in the first microepidemic of multidrug resistant tuberculosis recorded in the Czech Republic. Their disseminators were 21 individuals living in--or roaming between three distant areas. The age of 17 males ranged from 36 to 64 years (average 45 years) and of 4 females aged from 38 to 52 years. The index person was most probably a former male prisoner, aged 49 years, who disseminated multidrug resistant M. tuberculosis over a period of 28 months. In ten of the patients the following risk factors for tuberculosis were found: imprisonment, homelessness, immigration and previous stay in asylum--or in a psychiatric ward. In six cases, M. kansasii infection preceded tuberculosis. Four out of the 21 patients died. The RFLP analysis separated the patients into two distinct groups: group A comprising 14 members of which M. tuberculosis strains were isolated with six IS6110 copies, whereas the isolates of seven individuals of the group B, the RFLP profile displayed highly similar RFLP patterns compared to the isolates of group A, but with two additional IS6110 copies. In one patient, both A and B patterns were found: the first one in a M. tuberculosis strain isolated in 1993 and the second one in the isolate isolated two years later. Both the appearance of pattern B among the isolates of a part of patients and the switch from A to B pattern in one of patients can be plausibly explained by the unstability of DNA genotypes caused by transposition of IS6110 elements.

Adult↗
PubMed1999

A multidrug-resistant tuberculosis microepidemic caused by genetically closely related Mycobacterium tuberculosis strains.

IS6110 DNA fingerprinting was used to characterize an outbreak of multidrug-resistant tuberculosis in 21 individuals (17 males and 4 females) living in or roaming among four distantly separated areas in the Czech Republic. The restriction fragment length polymorphism (RFLP) analysis separated the collected Mycobacterium tuberculosis strains into group A, including 14 patients with six IS6110 copies, and group B, with 7 patients displaying highly similar RFLP patterns but with two additional IS6110 bands. A switch from pattern A to pattern B was observed in one patient, and the subsequent detection of subclone B in seven more individuals has been explained by the instability of DNA genotypes caused by transposition of IS6110 elements.

Czech Republic↗
PubMed1999

[A micro-epidemic of tuberculosis in the South Moravia Region].

In the South Moravian region which is the area with the lowest prevalence of bacillary tuberculosis in the Czech Republic (6.6 per 100,000 population in 1996) in 1993-1996 a total of six local and family microepidemics of tuberculosis were detected. For their identification the RFLP fingerprinting method was used based on evidence of repeated sequence of IS6110 in the chromosomal DNA of the examined strains of Mycobacterium tuberculosis predigested with restrictive enzyme PvuII. The profiles of fingerprints of each microepidemic were included by means of a computer programme into the hierarchy of the fingerprint dendrogram of 184 strains of M. tuberculosis which made it possible to identify possible identical profiles of strains from patients from remote places in the Czech Republic. In three family microepidemics involving always two members no identical fingerprint profiles of other Czech strains of M. tuberculosis were revealed. To the fourth cluster formed by six members of one family an identical RFLP profile of a female patient living in a nearby locality was added. In another microepidemic recorded in three brothers identical fingerprints were found another four patients from the South Moravian region and in one from the Central Bohemian region. The last cluster of two brothers was surprisingly enlarged by six identical RFLP profiles of patients from the West Bohemian region and one from Prague. These findings suggest that in areas with a low prevalence tuberculosis persists more frequently than in areas with a high prevalence as familial or local microepidemics.

Adolescent↗
PubMed1999

Present state of tuberculosis in the Czech Republic and in central European and Baltic countries.

The Central Europe forms a buffer zone between the countries of the European West reporting tuberculosis notification rates lower than 20 per 100,000, the cut-off set between low and high incidence areas, and the Eastern European countries including the republics of the former USSR, Russia and the Baltic States. The Czech Republic holds an intermediate place between these two territories with the total notification rate of tuberculosis cases 18.8, 9.7 bacteriologically verified and 5.7 positive in direct smear per 100,000 in 1996. Data on drug resistance obtained from the WHO/IUATLD Global Project on Anti-Tuberculosis Drug Resistance Surveillance were available from the Czech Republic, the only Central European country participating in the Project. The prevalence of resistant cases was here low: 2% primary and 13% acquired, and MDR cases were recorded in 1% of untreated and in 6% of repeatedly treated patients. The first microepidemic of MDR cases comprising 21 individuals was characterized by DNA fingerprinting. This outbreak pointed out the MDR tuberculosis as a new, extremely serious phenomenon in the epidemiology of tuberculosis. Corresponding data from Estonia and Latvia showed incomparably higher values in the drug resistance pattern: from 28 to 34% primary and 46 to 74% acquired resistance. MDR strains were reported in 9 to 14% of untreated and in 19 to 54% of repeatedly treated patients.

Adolescent↗
PubMed1999

Selected results of the tuberculosis control program in the Czech Republic.

There are probably several causes why steady decrease of tuberculosis incidence stopped. The aim of our work was to investigate a possible relation of tuberculosis incidence in smaller administrative areas with several social and economic characteristics. The individual data were taken from the Information System of Bacillary Tuberculosis, based on laboratory reporting network. Tuberculosis incidence dropped to 19.8/100,000 in 1987 and since then it fluctuates around that level. The levels in districts ranged from 1.9 to 45.6 in 1997. The social status, overcrowding, air pollution and unemployment rates are weakly correlated with tuberculosis incidence. These unfavourable values of social, economical and ecological indicators are linked with more industrialised parts of the country. Age and gender analysis shows that male population aged from 40-60 is the most affected population with higher rates of smear positive pulmonary TB. These rates are positively linked with a size of the municipality, in larger towns higher number of those cases are detected. Contrary to that, relatively higher rates of pulmonary tuberculosis are in elderly women in smaller municipalities. There is a suspicion about important role of relative poverty in the epidemiology of tuberculosis in the Czech Republic. The results indicate that we should concentrate our effort to reduce the incidence of tuberculosis mainly in economically active male population.

Adolescent↗
PubMed1997

[Serodiagnostic detection of IGA-specific antibodies in tuberculosis].

The objective of the submitted paper is evaluation of the possible use of detection of serum antibodies class IgA against the mycobacterial antigen Kp-90 ImCRA in the serodiagnostics of tuberculosis. Antibodies were detected by means of a commercially available IgA set Enzymimmunoassay Kreatech (Kreatech Diagnostics, Amsterdam, Netherlands). By means of the mentioned diagnostic kit antibodies class IgA were assessed in 26 patients with respiratory TB positive on microscopic examination and cultivation, 35 patients with tuberculosis of the lungs or extrapulmonary TB confirmed on cultivation and in 48 subjects with various clinical diagnoses, where TB was not confirmed clinically nor by bacteriological examination. In 64.3% of the patients with TB positive microscopical and cultivation optic density values (OD) higher than 1.2 were recorded, in 57% of these patients OD was higher than 1.5. The reactivity of patients in the control group (subjects with non-specific respiratory diseases) was strikingly low and all subjects with the exception of two had OD values lower than 1.2. The sensitivity of the test was 46.0%, the specificity 95.8%. The assembled results confirmed the reactivity of the majority of TB patients to mycobacterial antigen Kp-90 in the class of IgA serum antibodies and a low seroreactivity of controls with non-tuberculous affections of the airways where only two cases of high titres from a total of 48 examinations were recorded. The evaluated test--IgA Immunoassay Kreatech--thus extends the possibilities to detect the immunoreactivity of the patient when assessing the diagnosis and during the further follow up of the tuberculous finding.

Antibodies, Bacterial↗
PubMed1997

[Fingerprinting of mycobacterial DNA in the molecular epidemiology of tuberculosis].

A group of 24 strains of Mycobacterium tuberculosis was analyzed in a pilot study using the RFLP technique based on detection of IS6110 in chromosomal DNA predigested with PvuII. All strains had 5-15 copies of IS6110 and the method was also used in subsequent studies: From a total number of 15 strains obtained from patients from six families and institutions resp. according to the identity of finger prints transmission of TB infection was proved in 11 instances and ruled out in the remaining four because of a different RFLP profile of the finger prints. In the investigation of prevalence of different types of RFLP 75 strains of M. tuberculosis isolated consecutively during a four-month period in 1995 from patients living in Prague were analyzed. In the group a sub-group of 22 strains in seven clusters was differentiated which were formed by two to six strains with identical or related RFLP profiles and a group of 53 non-clustered strains with individually different RFLP patterns. The authors conclude from the assembled results that the great majority of findings corresponding to a 29% ratio of clustered strains is formed by endogenous reactivation from persistent tuberculous foci and the remaining part of 71% non-clustered strains is ascribed to recent exogenous superinfection.

Adult↗
PubMed1996

[Tuberculosis semper viva].

At the end of the twentieth century tuberculosis still has the attributes of a multiple, difficult to diagnose, and frequently treacherous disease which is difficult to treat. Due to its high prevalence in the developing countries and due to new medical and sociological elements associated with tuberculosis in economically developed countries with a low prevalence, tuberculosis still remains a paradoxical memento of medical science and practice. We know its etiological agent, treatment and principles of its control and despite this we must assume that it will persist in the human population for at least the period of one human life.

Czech Republic↗
PubMed1996

Bacillary tuberculosis in the Czech Republic: developmental trends in the 1981-1993 period.

Results of an automated information system on bacillary tuberculosis and mycobacterioses (ISBT) operating in the Czech Republic since 1981 nation-wide have been employed in this study. This system collects and processes data reported by all mycobacteriology laboratories in the country (34 in 1993) on each person disseminating pathogenic and/or opportunistic mycobacteria, on pathological materials examined in these patients and on methods applied for detection and identification of isolated mycobacteria Results of the 1981-1993 period were analyzed in this study. The annual incidence of bacillary tuberculosis cases identified by culture fell down from 2655 (25.8 per 100,000 popul.) in 1981 to 1139 (11.0 per 100,000 popul.) in 1993, i.e. by 57.4% in total, and by 4.8% in average annually. The decrease of annual mean values differed between two periods, from 1981 to 1985 and from 1986 to 1993, being 8.7% in the first and 3.7% in the successive period. The incidence of cases detected by direct microscopy of sputum showed a decline from 615 to 410 cases (5.97 to 3.97 per 100,000 popul.) in the 1981 to 1993 period, i.e. 2.8% annually. The analysis of the development and of the present state of the bacillary tuberculosis is instrumental in estimating the today's burden of the tuberculosis problem in the Czech Republic. Although distinct sings of worsened epidemiological parameters were not shown in this study, some disturbing findings can be considered as alerting: (a) a slowdown of the declining trend of bacillary tuberculosis cases detected by culture techniques seen in a few recent years, and (b) conserving potential tuberculosis pools in patients suffering from serious forms of the disease detectable by direct microscopy.

Czech Republic↗
PubMed1996

Use of IS6110 DNA fingerprinting in tracing man-to-man transmission of Mycobacterium tuberculosis in the Czech Republic.

The epidemiological relationship between tuberculosis cases in a prison and between cases within five families was investigated. Therefore, the isolated Mycobacterium tuberculosis strains were subjected to restriction fragment length polymorphism (RFLP) analysis using insertion sequence IS6110 as a probe. In case of 11 patients, the expected links of transmission were confirmed by RFLP typing. In contrast, in case of 4 patients the conclusion of classical contact tracing were not in agreement with the DNA fingerprinting results. These findings reinforce the usefulness of this recently developed technique as an additional tool in contact tracing. The IS6110DNA fingerprints of all strains investigated consisted of 7 to 13 bands and showed a high degree of polymorphism. Comparison of these fingerprints with those recorded in the Czech Republic previously, revealed the presence of a predominant DNA fingerprint type, without a known connection between the cases. Furthermore, other patterns found in the present study showed a high degree of similarity with the previously obtained fingerprints. Part of the patients were sampled twice. All of these double isolates showed identical fingerprints, confirming the previously described stability of IS6110DNA fingerprints. In contrast, from one couple of strains, isolated from husband and wife both suffering from tuberculosis, a slight change in one of the two patterns was observed. The patterns shared 10 band positions, confirming the expected relationship between these cases, but one of the patterns contained one additional band.

Contact Tracing↗