Publications by authors named "Giuseppe Stampone"

4 Publications

  • Page 1 of 1

Validation of a Commercial Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of spp. DNA in Processed Fish Products.

Foods 2020 Jan 16;9(1). Epub 2020 Jan 16.

Istituto Zooprofilattico Sperimentale della Sicilia, via Gino Marinuzzi 3, 90129 Palermo, Italy.

Parasites belonging to the genera are organisms of interest for human health because they are responsible for the Anisakiasis zoonosis, caused by the ingestion of raw or undercooked fish. Furthermore, several authors have reported this parasite to be a relevant inducer of acute or chronic allergic diseases. In this work, a rapid commercial system based on Loop-Mediated Isothermal Amplification (LAMP) was optimised and validated for the sensitive and rapid detection of spp. DNA in processed fish products. The specificity and sensitivity of the LAMP assay for processed fish samples experimentally infected with spp. larvae and DNA were determined. The LAMP system proposed in this study was able to give positive amplification for all the processed fish samples artificially contaminated with spp., giving sensitivity values equal to 100%. Specificity tests provided no amplification for the , , or genera and uninfected samples. The limit of detection (LOD) of the LAMP assay proposed was 10 times lower than the real-time PCR method compared. To the best of our knowledge, this is the first report regarding the application of the LAMP assay for the detection of spp. in processed fish products. The results obtained indicate that the LAMP assay validated in this work could be a reliable, easy-to-use, and convenient tool for the rapid detection of DNA in fish product inspection.
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http://dx.doi.org/10.3390/foods9010092DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7022600PMC
January 2020

Improvement of a rapid direct blood culture microbial identification protocol using MALDI-TOF MS and performance comparison with SepsiTyper kit.

J Microbiol Methods 2018 12 26;155:1-7. Epub 2018 Oct 26.

Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche, Università degli Studi di Palermo, Via Archirafi 32, I-90123 Palermo, Italy.

Fast diagnosis of pathogens is critical to guarantee the most adequate therapy for infections; bacterial culture methods, which constitute the actual gold standard, are precise and sensitive but rather slow. Today, new methods have been made available to enable faster diagnosis, with the Matrix-Assisted Laser Desorption Ionization-Time Of Flight Mass Spectrometry (MALDI-TOF MS) technique being the most promising. Even if simpler and faster than traditional bacterial culture methods, analysis of positive blood cultures via MALDI-TOF MS requires a preliminary extraction process of samples. In this study, we compared two extraction protocols for bacterial identification directly from positive blood cultures using the Bruker MALDI Biotyper system (Bruker Daltonics, Billerica, MA). In particular, we evaluated the time employed and the overall performance for their accurate identification. In this work, the performances of a commercial extraction kit, named SepsiTyper™ Kit, and those of the protocol developed by Treibmann et al. were evaluated and proven to be similar. However, the SELTERS method represents the best compromise price/performance. Lastly, an in-house developed analysis protocol has been tested, and the introduced optimizations granted a performance level equal if not better than the SepsiTyper kit, a reduced processing time and reduced costs.
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http://dx.doi.org/10.1016/j.mimet.2018.10.015DOI Listing
December 2018

Evaluation of a loop-mediated isothermal amplification method for the detection of in dairy food.

Ital J Food Saf 2017 Oct 14;6(4):6890. Epub 2017 Dec 14.

Department of Health, Animal Science and Food Safety, University of Milan.

Objective of the present study was to test the performances of a loop-mediated isothermal amplification (LAMP)-based method for the detection of , with particular focus on the dairy products. The specificity of the method was evaluated on 42 different spp. strains from collections, food and environmental samples. 100% (32 of 32) of the strains were correctly recognised, and none of other 10 spp. strains was misidentified. The sensitivity was evaluated on four strains from different sources. The instrument was able to detect 10-400 CFU/mL. The ability to detect low initial numbers of (0.3-0.7 Log CFU/g) was also evaluated, in duplicate, in pasteurised milk (whole and skimmed) and dairy samples (fresh ricotta, crescenza, mascarpone, mozzarella, cottage cheese, cream cheese, taleggio, gorgonzola). The analysis was performed after 18, 24 and 48 h of incubation, and was coupled with the count of in the broth. Microbial loads were insufficient to achieve a positive result after 18 and 24 h in most of the samples; after 48 h, all the products, except taleggio and one gorgonzola sample, were identified as positive; the sensitivity of the method when applied to contaminated dairy foods was about 5 Log CFU/g. The LAMP method tested can be considered a very useful tool, as it is a costeffective and easy-functioning method. The preliminary data obtained should be confirmed with a validation process taking into account different food typologies.
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http://dx.doi.org/10.4081/ijfs.2017.6890DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5850052PMC
October 2017

Identification of embryo-fetal cells in celomic fluid using morphological and short-tandem repeats analysis.

Prenat Diagn 2016 Oct 27;36(10):973-978. Epub 2016 Sep 27.

Unit of Hematology for Rare Diseases of Blood and Blood-forming Organs, Regional Reference Laboratory for Screening Prenatal Diagnosis of Hemoglobinopathies, Palermo, Italy.

Objective: The main problem to wide acceptability of celocentesis as earlier prenatal diagnosis is contamination of the sample by maternal cells. The objective of this study was to investigate the cellular composition of celomic fluid for morphological discrimination between maternal and embryo-fetal cells.

Method: Celomic fluids were aspired by ultrasound-guided transcervical celocentesis at 7-9 weeks' gestation from singleton pregnancies before surgical termination for psychological reasons. DNA extracted from celomic fluid cells showed the same morphology, and quantitative fluorescent polymerase chain reaction (PCR) assay was performed to evaluate their fetal or maternal origin.

Results: Six different types of non-hematological maternal and four different types of embryo-fetal cells were detected. The most common maternal cells were of epithelial origin. The majority of embryo-fetal cells were roundish with a nucleus located in an eccentric position near the wall. These cells were considered to be erythroblasts, probably derived from the yolk sac that serves as the initial site of erythropoiesis.

Conclusions: The combined use of morphology and DNA analysis makes it possible to select and isolate embryo-fetal cells, even when maternal contamination is high. This development provides the opportunity for the use of celocentesis for early prenatal diagnosis of genetic diseases and application of array comparative genomic hybridization. © 2016 John Wiley & Sons, Ltd.
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http://dx.doi.org/10.1002/pd.4922DOI Listing
October 2016
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