Search results for "Pcr assay"

Comparison of the artus Epstein-Barr virus (EBV) PCR kit and the Abbott RealTime EBV assay for measuring plasma EBV DNA loads in allogeneic stem cell…

2017

The ability of the artus Epstein-Barr virus (EBV) PCR kit and the Abbott RealTime EBV PCR assay to detect and quantify plasma EBV DNAemia was compared. The agreement between these assays was 95.8%. The EBV DNA loads measured by the two assays significantly correlated (P=< 0.0001).

Assessment of prevalence and load of torquetenovirus viraemia in a large cohort of healthy blood donors.

2020

OBJECTIVES: Torquetenovirus (TTV) is an emerging marker of functional immune competence with the potential to predict transplant-related adverse events. A large-scale epidemiological study was performed to understand how basal values vary in healthy individuals according to age and gender.; METHODS: We tested plasma from 1017 healthy blood donors aged 18-69years. The presence and load of TTV were determined by a real-time PCR assay. A sub-cohort of 384 donors was tested for anti-cytomegalovirus IgG antibodies, and 100 participants were also tested for TTV viraemia on a paired whole blood sample.; RESULTS: The overall prevalence of TTV was 65% (657/1017) with a mean (±SD) growth of 5±4% ever…

Field performance of the Abbott RealTime MTB assay for the diagnosis of extrapulmonary tuberculosis in a low-prevalence setting

2020

Introduction: The sensitivities of conventional mycobacterial culture in solid or liquid media and acid-fast bacilli (AFB) smear microscopy for Mycobacterium tuberculosis complex (MTBC) detection in extrapulmonary specimens are suboptimal. We evaluated the field performance of the Abbott RealTime MTB assay for the diagnosis of extrapulmonary tuberculosis in a low-prevalence setting. Methods: The total number of extrapulmonary specimens with mycobacterial culture and PCR results was 566: sterile fluids (n = 278), non-sterile fluids (n = 147), lymph node material (n = 69) tissue biopsies (n = 63), and abscess aspirates (n = 9). A composite standard consisting of mycobacterial culture results,…

Ubiquitous giants: a plethora of giant viruses found in Brazil and Antarctica

2018

Background Since the discovery of giant viruses infecting amoebae in 2003, many dogmas of virology have been revised and the search for these viruses has been intensified. Over the last few years, several new groups of these viruses have been discovered in various types of samples and environments.In this work, we describe the isolation of 68 giant viruses of amoeba obtained from environmental samples from Brazil and Antarctica. Methods Isolated viruses were identified by hemacolor staining, PCR assays and electron microscopy (scanning and/or transmission). Results A total of 64 viruses belonging to the Mimiviridae family were isolated (26 from lineage A, 13 from lineage B, 2 from lineage C…

Validation and implementation of a commercial real-time PCR assay for direct detection of Candida auris from surveillance samples.

2021

Background Rapid and reliable laboratory methods are required for detecting the nosocomial yeast Candida auris. AurisID® (Olm Diagnostics, England) is a real-time PCR assay approved for detecting C. auris in fungal cultures as well as directly from blood samples, involving a nucleic acid extraction as a prior step. Objectives The purpose of this study is to validate the AurisID® kit for direct detection of C. auris from surveillance samples without prior DNA extraction and to analyze the results of implementing this methodology to our daily laboratory routine protocol for C. auris surveillance studies. Methods Our PCR method using the AurisID® kit was compared with our routine protocol, con…

2020

Droplet microfluidics is a technology that enables the production and manipulation of small volumes. In biosciences, the most popular application of this technology is Droplet Digital™ PCR (ddPCR™), where parallel nanoliter-scale PCR assays are used to provide a high sensitivity and specificity for DNA detection. However, the recovery of PCR products for downstream applications such as sequencing can be challenging due to the droplets' stability. Here we compared five methods for disrupting the droplets to recover DNA. We found that rapid freezing in liquid nitrogen results in a clear phase separation and recovery of up to 70% of the DNA content. Liquid nitrogen freezing can thus offer a s…

Pre-emptive antiviral therapy for active CMV infection in adult allo-SCT patients guided by plasma CMV DNAemia quantitation using a real-time PCR ass…

2013

Pre-emptive antiviral therapy for active CMV infection in adult allo-SCT patients guided by plasma CMV DNAemia quantitation using a real-time PCR assay: clinical experience at a single center

Development of a TaqMan PCR assay for the identification of the non-native copepod Acartia tonsa, and detection of this species in Norwegian coastal …

2021

Abstract Molecular based assays for detection of species are a powerful tool to supplement morphological methods that may be time and labor intensive. Here we describe a sensitive TaqMan real time polymerase chain reaction assay that specifically detects the presence of Acartia tonsa in mixed plankton samples. The assay is used to find this non-native copepod in samples collected in Norwegian coastal waters.

Real-time PCR detection of Ochroconis lascauxensis involved in the formation of black stains in the Lascaux Cave, France

2012

A real-time Polymerase Chain Reaction (PCR) assay was developed to detect and quantify Ochroconis lascauxensis in the Lascaux Cave in France. This fungus is the principal causal agent of the black stains threatening the Paleolithic paintings of this UNESCO World Heritage Site. The black stains outbreak could not be stopped in spite of using intensive biocide treatments. A sensitive and time-saving protocol is needed for determining the extent of the colonization. Sets of primers that target the ITS and RPB2 regions were designed and evaluated for specificity against O. lascauxensis. Genomic DNA extracted from five species of Ochroconis and 13 other fungal species frequently isolated from ca…

Contamination of Wheat, Barley, and Maize Seeds with Toxigenic Fusarium Species and Their Mycotoxins in Tunisia

2021

Abstract Background Fusarium is a worldwide distributed fungal genus. It includes different species pathogenic to cereals among others crops. Some of these species can also produce toxic compounds toward animals and humans. Objective In this work, occurrence of fumonisins B1+B2, zearalenone, type A trichothecenes (T-2 and HT-2 toxins), and type B trichothecenes (deoxynivalenol[DON] and nivalenol[NIV]) was studied in 65 samples of stored and freshly harvested wheat, barley, and maize collected in Tunisia. Methods Mycotoxins analyses were performed by using gas chromatography for type B trichothecenes and HPLC for other mycotoxins. Obtained results were compared with the presence of mycotoxig…