Center for Cardiovascular Surgery and Transplantation Brno

Brno, Czech Republic

Center for Cardiovascular Surgery and Transplantation Brno

Brno, Czech Republic
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Nemcova E.,Center for Cardiovascular Surgery and Transplantation Brno | Nemcova E.,St Annes University Hospital Brno | Cernochova M.,Center for Cardiovascular Surgery and Transplantation Brno | Cernochova M.,Masaryk University | And 6 more authors.
PLoS ONE | Year: 2015

An increasing trend in non albicans infections and various susceptibility patterns to antifungal agents implies a requirement for the quick and reliable identification of a number of medically important Candida species. Real-time PCR followed by high resolution melting analysis (HRMA) was developed, tested on 25 reference Candida collection strains and validated on an additional 143 clinical isolates in this study. All reference strains and clinical isolates inconclusive when using phenotypic methods and/or HRMA were analysed using ITS2 sequencing. Considering reference and clinical strains together, 23 out of 27 Candida species could be clearly distinguished by HRMA, while the remaining 4 species were grouped in 2 pairs, when applying the mean Tm ± 3 SD values, the shape of the derivative melting curve (dMelt curve) and, in some cases, the normalized and temperature-shifted difference plot against C. krusei. HRMA as a simple, rapid and inexpensive tool was shown to be useful in identifying a wide spectrum of clinically important Candida species. It may complement the current clinical diagnostic approach based on commercially available biochemical kits. © 2015 Nemcova et al.


Ticha O.,Masaryk University | Stouracova M.,Masaryk University | Kuman M.,Center for Cardiovascular Surgery and Transplantation Brno | Studenik P.,Center for Cardiovascular Surgery and Transplantation Brno | And 2 more authors.
Transplant Immunology | Year: 2010

Background: Cytomegalovirus (CMV) infection is a life-threatening complication after solid organ transplantation. It usually appears in the first months after transplantation as a consequence of immunosuppression. The goal of this study was to evaluate the clinical significance of CD38high/CD3+8+ percentages in the detection of CMV infection in patients after kidney transplantation. Methods: In this retrospective study, 269 patients were monitored 2-3months after renal transplantation for the percentage of CD38high/CD3+8+ lymphocytes estimated by flow cytometry and for the number of CMV DNA copies in peripheral blood using a real-time polymerase chain reaction. Results: CMV infection was diagnosed in 12 (4.5%) patients between the 31st and 63rddays after transplantation, and all of them had percentages of CD38high/CD3+8+ T lymphocytes above 20%. In 4 of them, CMV DNAemia in peripheral blood was not detected, and 2 of these suffered from tissue-invasive CMV disease. In 7 patients with CMV DNAemia, the CD38high/CD3+8+ T lymphocyte percentage did not exceed 20%, and these patients did not develop CMV infection requiring antiviral treatment. In 23 additional patients, a CD38high/CD3+CD8+ percentage above 20% was recorded without CMV DNAemia. All of the remaining 234 patients never exceeded the arbitrary limit of 20%.The estimated sensitivity and specificity were 100% and 91% using clinical decision on the presence of CMV infection as a reference value, respectively. The estimated negative predictive value was 100%; however, the estimated positive predictive value was quite low (34%). Conclusions: The CD38high/CD3+8+ lymphocyte percentage seems to be a useful additional diagnostic marker for CMV infection in patients after kidney transplantation, especially when patients are in the risk of a tissue-invasive disease when CMV DNA copies may not be detectable in peripheral blood. © 2010 Elsevier B.V.


Vanerkova M.,Center for Cardiovascular Surgery and Transplantation Brno | Zaloudikova B.,Center for Cardiovascular Surgery and Transplantation Brno | Nemcova E.,Center for Cardiovascular Surgery and Transplantation Brno | Jurankova J.,University Hospital Brno | And 7 more authors.
Journal of Medical Microbiology | Year: 2010

Cardiobacterium valvarum, a fastidious Gram-negative bacterium, was detected in the aortic valve of a previously healthy 63-year-old man by broad-range PCR and 16S rRNA gene sequencing. In contrast to the patients in five previously published cases, our patient had neither a congenital bicuspid nor a prosthetic aortic valve. Here, we present a case of C. valvarum native tricuspid aortic valve infective endocarditis and a review of the literature. © 2010 SGM.


Obrucova H.,Center for Cardiovascular Surgery and Transplantation Brno | Obrucova H.,Masaryk University | Tihelkova R.,Center for Cardiovascular Surgery and Transplantation Brno | Kotaskova I.,Center for Cardiovascular Surgery and Transplantation Brno | And 7 more authors.
Journal of Clinical Microbiology | Year: 2016

Early diagnosis of fungal infection is critical for initiating antifungal therapy and reducing the high mortality rate in immunocompromised patients. In this study, we focused on rapid and sensitive identification of clinically important Candida species, utilizing the variability in the length of the ITS2 rRNA gene and fluorescent capillary electrophoresis (f-ITS2-PCR-CE). The method was developed and optimized on 29 various Candida reference strains from which 26 Candida species were clearly identified, while Candida guilliermondii, C. fermentati, and C. Carpophila, which are closely related, could not be distinguished. The method was subsequently validated on 143 blinded monofungal clinical isolates (comprising 26 species) and was able to identify 88% of species unambiguously. This indicated a higher resolution power than the classical phenotypic approach which correctly identified 73%. Finally, the culture-independent potential of this technique was addressed by the analysis of 55 retrospective DNA samples extracted directly from clinical material. The method showed 100% sensitivity and specificity compared to those of the combined results of cultivation and panfungal PCR followed by sequencing used as a gold standard. In conclusion, this newly developed f-ITS2-PCR-CE analytical approach was shown to be a fast, sensitive, and highly reproducible tool for both culture- dependent and culture-independent identification of clinically important Candida strains, including species of the "psilosis" complex. Copyright © 2016, American Society for Microbiology. All Rights Reserved.


PubMed | Center for Cardiovascular Surgery and Transplantation Brno
Type: Case Reports | Journal: Journal of medical microbiology | Year: 2010

Cardiobacterium valvarum, a fastidious Gram-negative bacterium, was detected in the aortic valve of a previously healthy 63-year-old man by broad-range PCR and 16S rRNA gene sequencing. In contrast to the patients in five previously published cases, our patient had neither a congenital bicuspid nor a prosthetic aortic valve. Here, we present a case of C. valvarum native tricuspid aortic valve infective endocarditis and a review of the literature.

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