For the first time microorganisms in CF sinuses are investigated by molecular methods in response to an absence of anaerobes in CF sinus samples during a two-year period at the Copenhagen CF center.
Endoscopic sinus surgery was performed in 19 CF patients. DNA from intact bacterial cells was investigated by 16S rRNA gene analysis and quantitative PCR. Results were compared to culture-dependent routine diagnosis.
Molecular methods showed a large microbial diversity, which included undetected anaerobes that may play a pathogenic role. Importantly, the culture methods did not always detect known CF pathogens. Quantitative PCR showed generally a higher abundance of classic CF pathogens e.g. Pseudomonas aeruginosa and Staphylococcus aureus compared with the anaerobe Propionibacterium acnes.
The results indicate that the culture methods in some cases may not be suitable as stand-alone method for this patient group, as diversity may be underestimated and important species undetected.
Escherichia coli strains from outpatient urinary tract infections in northern Norway over a period of 1 year were examined for resistance to nine commonly used antibiotics. Strains collected during 4.5 months were examined for R plasmid content by using conjugation and in vitro transformation. Of the E. coli strains, 42% were resistant to one or more antibiotics. Resistance was highest to sulfonamide (20.8% of all strains), nitrofurantoin (14.5%), and tetracycline (10.1%), whereas less than 6% of the strains were resistant to ampicillin, carbenicillin, cephalothin, nalidixic acid, or trimethoprim-sulfamethoxazole. No strain was resistant to gentamicin. Tetracycline resistance was more common in men than in women. Resistance to cephalothin, nalidixic acid, and sulfonamide was higher in strains from older people. Resistance to sulfonamide was more frequent in the urban community. These was no seasonal variation in antibiotic resistance, although the incidence of urinary tract infection varied with seasons. Plasmid-determined resistance to ampicillin, streptomycin, sulfonamide, and tetracycline was found. About 18% of the resistant strains from the urban municipality carried R plasmids, most of which were small plasmids mediating resistance to sulfonamide and streptomycin. The overall frequency of resistance in strains collected from rural areas was similar to the urban frequency, but in the rural strains, R plasmids were found in only 5% of the resistant strains.
OBJECTIVE: To study bacterial 16S RNA in archival prostate samples from 352 patients with benign prostate hyperplasia (BPH) and evaluate whether the presence of bacterial DNA was different in those who later developed prostate cancer (n = 171) and in the matched controls that did not progress to cancer (n = 181). METHODS: 16S DNA PCR followed by cloning and sequencing the positive samples. RESULTS: In 96/352 (27%) of the prostate tissue specimens 16S RNA were detected. Sequence analysis revealed Propionibacterium acnes as the predominant microorganism (23% of 16S RNA positive patients). The second most frequent isolate-Escherichia coli was found in 12 (12%) patients. The other isolates included Pseudomonas sp. (3 patients), Actinomyces sp. (2), Streptococcus mutans (1), Corynebacterium sp. (2), Nocardioides sp. (1), Rhodococcus sp. (1) Veillonella sp. (2). In P. acnes positive samples 62% exhibited severe histological inflammation versus 50% in the bacteria-negative group (p = 0.602). The presence of P. acnes in the prostate was associated with prostate cancer development (OR 2.17, 95% CI 0.77-6.95). CONCLUSIONS: This study has revealed P. acnes as the most common bacteria in the prostate in BPH. Further studies are needed to clarify its role in contributing to the development of prostatic inflammation and prostate cancer.
Cough is one of the most common complaints causing patients to seek medical attention, and chronic cough, defined as a cough period of at least three weeks, accounts for more than a third of referrals to a chest physician. Cough is an important factor in the spread and survival of microorganisms, but until recently little attention has been given to Bordetella pertussis (B. pertussis) in patients with chronic cough. This review summarizes the B. pertussis diagnostic methods--culture--polymerase chain reaction (PCR), and serology--and surveys the literature on B. pertussis and chronic cough in adults. There is growing evidence that B. pertussis is an important cause of persistent cough in adults; thus prevalence of pertusssis of 12.4-26% has been reported in studies from US, Australia and Germany. Recently we found evidence of pertussis as the cause of chronic cough in 16% of otherwise healthy adults in Denmark. Therefore, patients with chronic cough should be examined for B. pertussis infection. The demonstration of B. pertussis in an adult patient with chronic cough has two advantages: 1) the patient can be reassured that symptoms will disappear spontaneously, why more or less invasive examinations and empirical therapeutic trials can be omitted, 2) the source of infection can be eradicated, and contact persons, particularly non vaccinated infants in whom pertussis might be very severe, can be treated in order to avoid or attenuate clinical symptoms.
Infection with Brucella spp., long known as a cause of abortion, infertility, and reproductive loss in domestic livestock, has increasingly been documented in marine mammals over the past two decades. We report molecular evidence of Brucella infection in Asian sea otters (Enhydra lutris lutris). Brucella DNA was detected in 3 of 78 (4%) rectal swab samples collected between 2004 and 2006 on Bering Island, Russia. These 78 animals had previously been documented to have a Brucella seroprevalence of 28%, markedly higher than the prevalence documented in sea otters (Enhydra lutris) in North America. All of the DNA sequences amplified were identical to one or more previously isolated Brucella spp. including strains from both terrestrial and marine hosts. Phylogenetic analysis of this sequence suggested that one animal was shedding Brucella spp. DNA with a sequence matching a Brucella abortus strain, whereas two animals yielded a sequence matching a group of strains including isolates classified as Brucella pinnipedialis and Brucella melitensis. Our results highlight the diversity of Brucella spp. within a single sea otter population.
The study objective was to get more information on C. burnetii prevalence in wild birds and ticks feeding on them, and the potentialities of the pathogen dissemination over Europe by both.
Blood, blood sera, feces of wild birds and ticks removed from those birds or from vegetation were studied at two sites in Russia: the Curonian Spit (site KK), and the vicinity of St. Petersburg (site SPb), and at two sites in Bulgaria: the Atanasovsko Lake (site AL), and the vicinity of Sofia (site SR).
C. burnetii DNA was detected in blood, feces, and ticks by PCR (polymerase chain reaction). All positive results were confirmed by Sanger's sequencing of 16SrRNA gene target fragments. The antibodies to C. burnetii in sera were detected by CFR (complement fixation reaction).
Eleven of 55 bird species captured at KK site hosted Ixodes ricinus. C. burnetii DNA was detected in three I. ricinus nymphs removed from one bird (Erithacus rubecula), and in adult ticks flagged from vegetation: 0.7% I. persulcatus (site SPb), 0.9% I. ricinus (site KK), 1.0% D. reticulatus (AL site). C. burnetii DNA was also detected in 1.4% of bird blood samples at SPb site, and in 0.5% of those at AL site. Antibodies to C. burnetii were found in 8.1% of bird sera (site SPb). C. burnetii DNA was revealed in feces of birds: 0.6% at AL site, and 13.7% at SR site.
Both molecular-genetic and immunological methods were applied to confirm the role of birds as a natural reservoir of C. burnetii. The places of wild bird stopover in Russia (Baltic region) and in Bulgaria (Atanasovsko Lake and Sofia region) proved to be natural foci of C. burnetii infection. Migratory birds are likely to act as efficient "vehicles" in dispersal of C. burnetii -infested ixodid ticks.
To study specific features of the incidence, course and diagnosis of tuberculosis pericarditis (TP) in patients with HIV-infection.
We analysed results of diagnosis of 304 primary patients with organ tuberculosis in combination with HIV infection treated in Moscow tuberculosis hospital N 7 in 2006-2010. CD4 lymphocyte count median in tuberculosis onset was 140 in 1 mcl, 63.2% patients had a baseline level of CD4 lymphocytes under 200 in 1 mcl.
TP incidence in primary patients with tuberculosis and HIV-infection was 6.3% while in patients with tuberculosis of multiple locations--13.7%. Cardiac tamponade symptoms were registered only in one case. Pericardial effusion was classified as moderate and large in 68.4% patients. Patients with large effusion (more than 20 mm in isolation of pericardial leaves) have undergone diagnostic pericardiocentesis and, in some cases, microdrainage. Sensitivity of exudate test for M. tuberculosis DNA with use of polymerase chain reaction was 100%.
Active surgical policy in massive effusion tuberculosis pericarditis in line with adequate antituberculosis and antiretrovirus therapy in HIV-infected patients results in rapid resorption of the effusion.
An outbreak of pertussis in Manitoba, Canada, provided an opportunity to evaluate the recently developed monoclonal antibody (MAb) BL-5 for the direct detection of Bordetella pertussis. The MAb recognizes a lipooligosaccharide epitope. A total of 1,507 consecutive nasopharyngeal swabs for culture and companion smears for direct fluorescent-antibody (DFA) detection were evaluated at Cadham Provincial Laboratory between September and November 1994. The cutoff for DFA positivity was four fluorescing organisms with morphology characteristic of B. pertussis. PCR analysis for B. pertussis DNA was performed on a subset of 100 smears by eluting material from the slides after DFA examination. In comparison with culture, the sensitivity, specificity, and positive and negative predictive values of BL-5 were 65.1% (41 of 63 samples), 99.6% (1,438 of 1,444 samples), 87.2% (41 of 47 samples), and 98.5% (1,438 of 1,460 samples), respectively. The sensitivity of culture compared with PCR was 45.5% (10 of 22 samples) for the subset of 100 specimens tested by both procedures. An expanded "gold standard" of positivity by culture or PCR for these 100 specimens resulted in DFA sensitivity, specificity, and positive and negative predictive values of 32.3, 97.1, 83.3, and 76.1%, respectively. The utility of MAb BL-5 for direct detection of B. pertussis in a clinical laboratory setting has been demonstrated by this investigation.
Improvement of monitoring and prognosis of epidemic manifestations of natural foci of tularemia on the territory of Voronezh region using immune-serological and molecular-genetic study of main carriers of the disease.
539 small mammals captured during summer period of 2011 in 4 districts of North-Eastern part of Voronezh region were studied. Animal organs were studied by serologic (search for Francisella tularensis antigens) and molecular-biologic (detection of F. tularensis DNA) methods. Tularemia antigen was detected using passive hemagglutination reaction (PHAR) with erythrocytic tularemia immunoglobulin diagnosticum. Real-time polymerase chain reaction (RT-PCR) was applied for detection of tularemia causative agent DNA.
Complex study revealed epizootic activity of natural foci of tularemia in the examined territory. F. tularensis antigen and/or DNA were detected in 82 objects (15.2%). Use of RT-PCR allowed to additionally detect samples with relatively low content of F. tularensis DNA substrate, when antigen was not detected in samples. High sensitivity and specificity of the RT-PCR was ensured by inclusion of specific probes (tu14-PR2 and ISFTu2P).
The results obtained give evidence on functioning and epizootic activity of natural foci of tularemia in Voronezh region that requires constant monitoring of the territory and prophylaxis measures, first of all vaccination of risk groups by live tularemia vaccine.
The association of Chlamydia pneumoniae with atherosclerosis is still controversial. Reports from different laboratories have varied widely and "gold standards" for the detection of C. pneumoniae are lacking. In the present study, aortic valves and peripheral blood mononuclear cells from 48 patients undergoing aortic valve replacement were examined for the presence of C. pneumoniae using a nested PCR. C. pneumoniae-specific DNA was not detected in any of the clinical samples. No PCR inhibition was observed by spiking the samples with target C. pneumoniae. A total of 31/46 patients (67%) were seropositive for C. pneumoniae IgG. These results do not support the association of C. pneumoniae with aortic valves and peripheral blood mononuclear cells in patients with atherosclerotic aortic heart valve disease.