Search results for "real time PCR"

Cause and duration of mustard incorporation effects on soil-borne plant pathogenic fungi

2009

International audience; Two fungal plant pathogens, Rhizoctonia solani AG 2-2 and Fusarium oxysporum f.sp. lini, were studied in relation to general responses of soil fungi and bacteria following incorporation of Brassica juncea. Our aim was to understand to what extent the changes in the biological and physicochemical characteristics of the soil could explain the effects on the studied pathogens and diseases, and to determine the temporal nature of the responses. Short-term effects of mustard incorporation (up to 4 months) were investigated in a microcosm experiment, and compared with a treatment where composted plant material was incorporated. In a field experiment, the responses were fol…

Design and Performance Testing of a DNA Extraction Assay for Sensitive and Reliable Quantification of Acetic Acid Bacteria Directly in Red Wine Using…

2016

International audience; Although strategies exist to prevent AAB contamination, the increased interest for wines with low sulfite addition leads to greater AAB spoilage. Hence, there is a real need for a rapid, specific, sensitive, and reliable method for detecting these spoilage bacteria. All these requirements are met by real time Polymerase Chain Reaction (or quantitative PCR; qPCR). Here, we compare existing methods of isolating DNA and their adaptation to a red wine matrix. Two different protocols for isolating DNA and three PCR mix compositions were tested to select the best method. The addition of insoluble polyvinylpolypyrrolidone (PVPP) at 1% (v/v) during DNA extraction using a pro…

Rapid, cost-effective, sensitive and quantitative detection of Acinetobacter baumannii from pneumonia patients

2011

Background and Objectives: Pneumonia with Acinetobacter baumannii has a major therapeutic problem in health care settings. Decision to initiate correct antibiotic therapy requires rapid identification and quantification of organism. The aim of this study was to develop a rapid and sensitive method for direct detection of A. baumannii from respiratory specimens. Materials and Methods: A Taqman real time PCR based on the sequence of blaoxa-51 was designed and used for direct detection of A. baumannii from 361 respiratory specimens of patients with pneumonia. All specimens were checked by conventional bacteriology in parallel. Results: The new real time PCR could detect less than 200 cfu per m…

Application of fnbA gene as new target for the species-specific and quantitative detection of Staphylococcus aureus directly from lower respiratory t…

2013

Staphylococcus aureus is a significant cause of hospital-acquired pneumonia (HAP), particularly in mechanically ventilated patients. We used the fibronectin-binding protein A gene (fnbA) for the species-specific and quantitative detection of S. aureus directly from lower respiratory tract (LRT) specimens by a Taq Man real time PCR. For this reason, a total of 269 lower respiratory tract (LRT) specimens collected from patients with hospital-acquired pneumonia were assayed. Amplification of fnbA in serial dilutions ranged from 10(9) CFU/ ml to 10(2) CFU/ml. Standard curve of triplicate every dilution had slope 3.34±0.1 and R2>0.99 with SD 0.1. Based on these data, the sensitivity and specif…

Development of methods for the detection and quantification of spoilage microorganisms in wine : study of growing factors

2016

New practices used to elaborate wine lead to an increase of wine spoilage due to microorganisms. That is why, new technics have to be developed to quantify these microorganisms accurately, quickly and with low costs. The main wine spoilages are due to acetic acid bacteria (AAB) (A. aceti, A. pasteurianus, G. oxydans and Ga. liquefaciens) and Brettanomyces bruxellensis development. AAB transforms ethanol to acetic acid while B. bruxellensis transforms hydroxycinnamic acids to ethyl phenols (EP) (unpleasant odor molecules). In order to detect these wine spoilage microrganisms, flow cytometry coupled to fluorescent in situ hybridization has been assessed. No reproducible results have been deve…

Polymerase Chain Reaction (PCR)

2008

AUTOPHAGY AND APOPTOSIS MODULATION BY AQUEOUS EXTRACTS FROM LEAVES AND RHIZOMES OF Posidonia oceanica ON HEPG2 HEPATOCARCINOMA CELLS

2023

Single tube real time PCR for detection of Streptococcus pneumoniae, Mycoplasma pneumoniae, Chlamydophila pneumoniae and Legionella pneumophila from …

2012

We designed a multiplex real time PCR for rapid, sensitive and specific detection of Streptococcus pneumoniae, Legionella pneumophila, Chlamydophila pneumoniae and Mycoplasma pneumoniae. The study cases consisted of 129 patients with community acquired pneumonia (CAP). Bacteriological techniques were implemented for detection of the cultivable organisms. DNA were extracted from sputa, throat swabs, bronchoalveolar lavages and tracheal aspirates and used as templates in real time PCR. The primers and probes were designed for cbpA (S. pneumoniae), p1adhesin (M. pneumoniae), mip (L. pneumophila) and ompA (C. pneumoniae). After optimization of real time PCR for every organism, the experiments w…

Coxiella burnetii spread in Sicily

2015

Q fever is a zoonotic disease caused by C. burnetii. Common reservoirs of this worldwide disease are wild and domestic animals, especially sheep, goats, cattle. Q fever has been considered also an occupational disease for abattoir workers, sheep shearers, livestock farmers, and veterinarians due to their direct contact with potentially infected animals. The aims of this study were to estimate C. burnetii spread in Sicilian livestock and among rangers that live and work in Western Sicily. ELISA test on animal serum and IFI test on seasonal rangers sera were carried out. Real Time PCR was performed on milk and vaginal swab samples collected from animals to search for the C. burnetii DNA. The …

Quantification and viability assays of Toxoplasma gondii in commercial "Serrano" ham samples using magnetic capture real-time qPCR and bioassay techn…

2014

"Serrano" ham is a typical pork product from the Mediterranean area, highly valued for its flavour. To make Serrano ham, pork undergoes a salting and a subsequent fermentation process known as curing. Certain pigs used for meat production are an important source of Toxoplasma gondii infection in humans. We have developed a method for quantifying and assaying the viability of the T. gondii present in commercial Serrano ham samples. A magnetic capture method for the isolation of T. gondii DNA and a qRT-PCR were used to estimate the T. gondii burden in 475 commercial samples of "Serrano" ham in two presentation formats: ham pieces and sliced ham. The infectivity capacity of T. gondii in positi…