scholarly journals Detection of Multiple Fungal Species in Blood Samples by Real-Time PCR: an Interpretative Challenge

2014 ◽  
Vol 52 (9) ◽  
pp. 3515-3516 ◽  
Author(s):  
Catriona L. Halliday ◽  
Tania C. Sorrell ◽  
Sharon C.-A. Chen
2014 ◽  
Vol 52 (9) ◽  
pp. 3517-3517
Author(s):  
Yanan Zhao ◽  
Dimitrios P. Kontoyiannis ◽  
Emmanuel Roilides ◽  
Thomas J. Walsh ◽  
David S. Perlin

2010 ◽  
Vol 4 (08) ◽  
pp. 511-516 ◽  
Author(s):  
Parisa Badiee ◽  
Abdolvahab Alborzi

Introduction:  Invasive aspergillosis is a severe complication of cytotoxic chemotherapies and bone marrow transplantation (BMT). The aim of this study was to assess the utility of a real-time PCR assay for the early diagnosis of Aspergillus species in blood samples from BMT patients. Methodology: Blood specimens (n = 993) from patients (n = 82) scheduled for BMT were collected prior to transplant and for 100 days post transplantation.  The specimens were later tested using an Aspergillus-specific real-time PCR assay. Cultures of clinical samples, along with sonography and computerized tomographic scans, were performed as standard of care. Results: Aspergillus DNA was positive in 94 sequential blood samples from 13 patients with clinical and radiological signs of infection. Samples from three of these patients were PCR-positive for Aspergillus in the first week of admission, prior to transplantation. Four patients with aspergillosis were cured with antifungal agents and nine died. An additional 12 patients without clinical signs of infection were PCR-positive on one occasion each, while two patients with clinical signs of infection were PCR-negative. Compared to routine methods of aspergillosis diagnosis, the respective sensitivity, specificity, negative, and positive predictive values of the PCR method by patient were 86.6%, 82%, 96.5% and 52%. Conclusions: The results show that Aspergillus infections in the blood of bone marrow transplant patients can be dectected by PCR methods. Early detection of Aspergillus infections by PCR has the potential to positively impact patient mortality rate and provide cost savings to hospitals.


2010 ◽  
Vol 24 (3) ◽  
pp. 134-138 ◽  
Author(s):  
Kenan Sener ◽  
Mehmet Yapar ◽  
Orhan Bedir ◽  
Cem Gül ◽  
Ömer Coskun ◽  
...  

2016 ◽  
Vol 60 (1) ◽  
pp. 7-12 ◽  
Author(s):  
Aliasghar Bahari ◽  
Masoud Sabouri Ghannad ◽  
Omid Dezfoulian ◽  
Fereydon Rezazadeh ◽  
Ali Sadeghi-Nasab

Abstract Introduction: The aim of this study was to use TaqMan real-time PCR technique to investigate Jaagsiekte sheep retrovirus (JSRV) proviral DNA in whole blood samples of sheep, and compare the results to those of histopathological examinations. Material and Methods: Eighty blood samples from clinically healthy sheep were randomly collected before the animals were slaughtered. Ten tissue samples from each lung and associated caudal mediastinal lymph node were taken. Results: Fifteen (18.75%) blood samples were found to contain proviral DNA, and 11 (13.75%) corresponding lung samples showed microscopic changes consistent with ovine pulmonary adenocarcinoma. None of the samples displayed metastases to the caudal mediastinal lymph nodes. The prominent pattern of neoplastic nodules consisted of acinar (alveolar) form. Conclusion: The results indicated the higher sensitivity of real-time PCR compared to histopathological examinations in detection of ovine pulmonary adenocarcinoma.


Author(s):  
Madeleine Mai ◽  
Iris Müller ◽  
Daniela Maneg ◽  
Benedikt Lohr ◽  
Achim Haecker ◽  
...  

2009 ◽  
Vol 58 (8) ◽  
pp. 1037-1044 ◽  
Author(s):  
Nobutoshi Soeta ◽  
Masanori Terashima ◽  
Mitsukazu Gotoh ◽  
Shuichi Mori ◽  
Kyoko Nishiyama ◽  
...  

To develop a rapid and quantitative diagnostic technique for the detection and identification of a wide range of fungi, an improved molecular method based on real-time PCR and the analysis of its products that targets the internal transcribed spacer (ITS) 2 region was established. The real-time PCR could quantitatively and specifically detect the ITS2 region from all 24 tested pathogenic fungal species at between 101 and 107 copies per test without amplification of bacterial or human DNA. The sequences of the primer-binding sites are conserved in the registered sequences of 34 other pathogenic fungal species, suggesting that the PCR would also detect these species. The hyperpolymorphic nature of the ITS2 region between fungal species in terms of length and nucleotide sequence provided valuable information for the determination of species. By labelling the 5′ end of the reverse primer with NED fluorescent dye, the fragment lengths of the real-time PCR products and their 3′-terminal fragments, derived using restriction enzyme ScrFI digestion, were easily evaluated by capillary electrophoresis. Using this analysis, the number and species of fungi present in samples could be estimated. Moreover, sequence analysis of the real-time PCR products could accurately determine species in samples containing a single species. This diagnostic technique can estimate a wide range of fungi from various clinical samples within 1 day and accurately identify them in 2 days. Quantitative results for fungal titre in samples can also provide useful information for understanding the progression of disease and the efficacy of antifungal chemotherapy.


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