Satellite DNA as a target for TaqMan real-time PCR detection of the pinewood nematode, Bursaphelenchus xylophilus

The pinewood nematode (PWN), Bursaphelenchus xylophilus , is a major pathogen of conifers, which impacts on forest health, natural ecosystem stability and international trade. As a consequence, it has been listed as a quarantine organism in Europe. A real-time PCR approach based on TaqMan chemistry was developed to detect this organism. Specific probe and primers were designed based on the sequence of the Msp I satellite DNA family previously characterized in the genome of the nematode. The method proved to be specific in tests with target DNA from PWN isolates from worldwide origin. From a practical point of view, detection limit was 1 pg of target DNA or one individual nematode. In addition, PWN genomic DNA or single individuals were positively detected in mixed samples in which B. xylophilius was associated with the closely related non-pathogenic species B. mucronatus , up to the limit of 0.01% or 1% of the mixture, respectively. The real-time PCR assay was also used in conjunction with a simple DNA extraction method to detect PWN directly in artificially infested wood samples. These results demonstrate the potential of this assay to provide rapid, accurate and sensitive molecular identification of the PWN in relation to pest risk assessment in the field and quarantine regulation.

A Case Based approach to assess Waiting Time Prediction at an Intensive Care Unity

Waiting time at an intensive care unity stands for a key feature in the assessment of healthcare quality. Nevertheless, its estimation is a difficult task, not only due to the different factors with intricate relations among them, but also with respect to the available data, which may be incomplete, self-contradictory or even unknown. However, its prediction not only improves the patients’ satisfaction but also enhance the quality of the healthcare being provided. To fulfill this goal, this work aims at the development of a decision support system that allows one to predict how long a patient should remain at an emergency unit, having into consideration all the remarks that were just stated above. It is built on top of a Logic Programming approach to knowledge representation and reasoning, complemented with a Case Base approach to computing.