A Valuation of the Reliability of a GIS Based on the Fuzzy Logic in a Concrete Case Study

The great difficulty of evaluating the quality of the applications concerning environmental problems, mainly for the heterogeneity of input data and their indeterminateness in the error estimations, is a well known problem. The usage of the Fuzzy Logic can be adequate in the treatment of this kind of information, especially when using approximate linguistic labels to define the input data. We have applied this idea in a previous work and here proposed for the study of a GIS of the PROCIDA island (located near Napoli), realized with technology of the Environmental Systems Research Institute and implemented by means of a software tool called FUZZY-SRA

Performance of various methods used for detecting WSSV DNA.

<p>Performance of various methods used for detecting WSSV DNA.</p

PCR primers used in this study.

<p>WSSV PCR-757 FP and RP were used for the PCR-cloning of <i>vp28</i>.</p><p>VP28–140F and R were used in qPCR assay.</p

Probit regression of WSSV real-time RPA using the data of 8 runs.

<p>(The sensitivity of 10 molecules with 95% reliability by SPSS probit regression analysis).</p

Analytical specificity of <i>Fno</i> RPA.

<p>Analytical specificity of <i>Fno</i> RPA.</p

Probit regression analysis of data set of 10 runs of <i>Fno</i>-RPA and qPCR using Minitab^®.

<p>The black triangle indicates limits of detection at 95% probability which were 15 and 11 molecules detected of <i>Fno</i>-plasmid standard DNA in RPA (A) and qPCR (B) respectively.</p

List of primers and probes used in the experiment.

<p>List of primers and probes used in the experiment.</p

Screening of tilapia tissues and water samples for <i>Fno</i>.

<p>Screening of tilapia tissues and water samples for <i>Fno</i>.</p

The diagnostic performance of the <i>Fno</i> RPA using field samples.

<p>The diagnostic performance of the <i>Fno</i> RPA using field samples.</p

Rapid Isothermal Detection of Pathogenic Clostridioides difficile Using Recombinase Polymerase Amplification

Nosocomial-associated diarrhea due to Clostridioides difficile infection (CDI) is diagnosed after sample precultivation by the detection of the toxins in enzyme immunoassays or via toxin gene nucleic acid amplification. Rapid and direct diagnosis is important for targeted treatment to prevent severe cases and recurrence. We developed two singleplex and a one-pot duplex fluorescent 15 min isothermal recombinase polymerase amplification (RPA) assays targeting the toxin genes A and B (tcdA and tcdB). Furthermore, we adapted the singleplex RPA to a 3D-printed microreactor device. Analytical sensitivity was determined using a DNA standard and DNA extracts of 20 C. difficile strains with different toxinotypes. Nineteen clostridial and gastrointestinal bacteria strains were used to determine analytical specificity. Adaptation of singleplex assays to duplex assays in a 50 μL volume required optimized primer and probe concentrations. A volume reduction by one-fourth (12.4 μL) was established for the 3D-printed microreactor. Mixing of RPA was confirmed as essential for optimal analytical sensitivity. Detection limits (LOD) ranging from 119 to 1411 DNA molecules detected were similar in the duplex tube format and in the singleplex 3D-printed microreactor format. The duplex RPA allows the simultaneous detection of both toxins important for the timely and reliable diagnosis of CDI. The 3D-printed reaction chamber can be developed into a microfluidic lab-on-a-chip system use at the point of care