scholarly journals Molecular Detection and Species-Specific Identification of Medically Important Aspergillus Species by Real-Time PCR in Experimental Invasive Pulmonary Aspergillosis

2011 ◽  
Vol 49 (12) ◽  
pp. 4150-4157 ◽  
Author(s):  
T. J. Walsh ◽  
M. C. Wissel ◽  
K. J. Grantham ◽  
R. Petraitiene ◽  
V. Petraitis ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Molecular Detection ◽  
Invasive Pulmonary Aspergillosis ◽  
Aspergillus Species ◽  
Pulmonary Aspergillosis ◽  
Specific Identification ◽  
Species Specific

scholarly journals Rapid Species Identification of Cooked Poisonous Mushrooms by Using Real-Time PCR

2008 ◽  
Vol 74 (10) ◽  
pp. 3306-3309 ◽  
Author(s):  
Kazuhiko Maeta ◽  
Tomoya Ochi ◽  
Keisuke Tokimoto ◽  
Norihiro Shimomura ◽  
Nitaro Maekawa ◽  
...  
Keyword(s):  
Real Time ◽  
Species Identification ◽  
Real Time Pcr ◽  
Specific Test ◽  
Specific Identification ◽  
Specific Fluorescence ◽  
Species Specific ◽  
Poisonous Mushrooms

ABSTRACT Species-specific identification of the major cooked and fresh poisonous mushrooms in Japan was performed using a real-time PCR system. Specific fluorescence signals were detected, and no nonspecific signals were detected. Therefore, we succeeded in developing a species-specific test for the identification of poisonous mushrooms within 1.5 h.

Species-specific identification of variola, monkeypox, cowpox, and vaccinia viruses by multiplex real-time PCR assay

2011 ◽  
Vol 175 (2) ◽  
pp. 163-169 ◽  
Author(s):  
Sergei N. Shchelkunov ◽  
Dmitrii N. Shcherbakov ◽  
Rinat A. Maksyutov ◽  
Elena V. Gavrilova
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Assay ◽  
Specific Identification ◽  
Vaccinia Viruses ◽  
Species Specific

scholarly journals Aspergillus-Specific Lateral-Flow Device and Real-Time PCR Testing of Bronchoalveolar Lavage Fluid: a Combination Biomarker Approach for Clinical Diagnosis of Invasive Pulmonary Aspergillosis

2015 ◽  
Vol 53 (7) ◽  
pp. 2103-2108 ◽  
Author(s):  
Gemma L. Johnson ◽  
Shah-Jalal Sarker ◽  
Francesco Nannini ◽  
Arianna Ferrini ◽  
Emma Taylor ◽  
...  
Keyword(s):  
Bronchoalveolar Lavage ◽  
Real Time ◽  
Antifungal Therapy ◽  
Real Time Pcr ◽  
Invasive Pulmonary Aspergillosis ◽  
Lateral Flow ◽  
Pulmonary Aspergillosis ◽  
Lateral Flow Device ◽  
Flow Device ◽  
The Impact

Clinical experience with the impact of serum biomarkers for invasive fungal disease (IFD) varies markedly in hemato-oncology. Invasive pulmonary aspergillosis (IPA) is the most common manifestation, so we evaluated biomarkers in bronchoalveolar lavage (BAL) fluid. AnAspergillus-specific lateral-flow device (LFD), quantitative real-time PCR (qPCR), and the galactomannan (GM) test were used with 32 BAL fluid samples from 32 patients at risk of IPA. Eight patients had proven IPA, 3 had probable IPA, 6 had possible IPA, and 15 patients had no IPA by European Organization for Research and Treatment of Cancer Invasive Fungal Infections Cooperative Group/Mycoses Study Group of the National Institute of Allergy and Infectious Diseases (EORTC/MSG) criteria. The diagnostic accuracies of the tests were evaluated, and pairwise agreement between biomarkers was calculated. The diagnostic performance of the EORTC/MSG criteria was evaluated against the test(s) identified to be the most useful for IPA diagnosis. Using the EORTC/MSG criteria, the sensitivities of qPCR and LFD were 100% and the sensitivity of the GM test was 87.5% (GM test index cutoff, >0.8), with the tests having specificities of between 66.7 and 86.7%. The agreement between the results of qPCR and LFD was almost perfect (Cohen's kappa coefficient = 0.93, 95% confidence interval, 0.81 to 1.00). LFD and qPCR combined had a sensitivity of 100% and a specificity of 85.7%. Calcofluor staining and culture of all BAL fluid samples were negative for fungal infection. The median time from the start of mold-active antifungal therapy to the time of collection of BAL fluid was 6 days. Reversing roles and using dual testing by LFD and qPCR to classify cases, the EORTC/MSG criteria had a sensitivity of 83.3%. All three tests are useful for the diagnosis of IPA in BAL fluid samples. Despite the significant delays between the start of antifungal therapy and bronchoscopy, unlike microscopy and culture, the biomarkers remained informative. In particular, the combination of LFD and qPCR allows the sensitive and specific detection of IPA.

Conventional and Real-Time PCR–Based Approaches for Molecular Detection and Quantitation of Bovine Species Material in Edible Gelatin

2005 ◽  
Vol 68 (11) ◽  
pp. 2420-2426 ◽  
Author(s):  
TAURAI TASARA ◽  
SANDRA SCHUMACHER ◽  
ROGER STEPHAN
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Molecular Detection ◽  
Raw Material ◽  
Spongiform Encephalopathy ◽  
Specific Pcr ◽  
Species Specific ◽  
Bovine Gelatin ◽  
Bovine Species ◽  
Pcr Primer

The majority of edible gelatin in Europe is derived from pigskin, but a significant portion is extracted from bovine tissue. Because of the bovine spongiform encephalopathy crisis, consumers might be concerned about the gelatin used in various products. To assure consumers of the quality and safety of edible gelatin, European Union directive 1999/724/EC described general guidelines for gelatin production, including requirements for documentary proof confirming that raw materials are from animals fit for human consumption. Analytical methods to confirm gelatin documentation or raw material animal species source in the finished product are lacking. In this study, several published species-specific PCR systems were evaluated as potential molecular methods for determining the origin of the raw material used in making gelatin. A recently validated bovine species-specific PCR primer set targeting the ATPase 8 subunit gene in bovine mitochondrial DNA was suitable for detection of bovine material in gelatin. This PCR primer set was optimized using conventional and real-time PCR approaches. An evaluation of these two PCR methods confirmed the high specificity for the adopted primer set in various gelatin matrices of known origin. The inclusion of bovine gelatin in pork or fish gelatin can be detected at 0.1 to 0.001%. These PCR assays are potential molecular detection tools that can be used to routinely detect bovine gelatin either alone or as an inclusion in gelatin made from other species.

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