C Vocale1, S G Rimoldi2, C Pagani2, R Grande2, F Pedna3, M Arghittu4, G Lunghi4, A Maraschini4, M R Gismondo2, M P Landini5, E Torresani4, F Topin6, V Sambri7. 1. Unit of Clinical Microbiology, Regional Reference Centre for Microbiological Emergencies, St. Orsola Malpighi University Hospital, Via Massarenti 9, 40138 Bologna, Italy. Electronic address: cate.vocale@gmail.com. 2. Laboratory of Clinical Microbiology, Virology and Bioemergency, "L. Sacco" University Hospital, Milan, Italy. 3. Unit of Clinical Microbiology, The Hub Laboratory of the Greater Romagna Area, Pievesestina, Cesena, Italy. 4. Department of Clinical Chemistry and Microbiology, Bacteriology and Virology Units, Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico, Milan, Italy. 5. Unit of Clinical Microbiology, Regional Reference Centre for Microbiological Emergencies, St. Orsola Malpighi University Hospital, Via Massarenti 9, 40138 Bologna, Italy; DIMES, University of Bologna, Bologna, Italy. 6. Luminex Corporation, Oosterhout NB, Netherlands. 7. Unit of Clinical Microbiology, The Hub Laboratory of the Greater Romagna Area, Pievesestina, Cesena, Italy; DIMES, University of Bologna, Bologna, Italy.
Abstract
OBJECTIVE: Gastroenteritis caused by a single pathogen or multiple pathogens remains a major diagnostic challenge for the laboratory. The treatment of diarrhoea is based on microbiological results. Diagnosis is achieved using different laboratory techniques that have variable sensitivity and specificity. xTAG GPP is a new multiplex PCR assay that simultaneously detects 15 different pathogens responsible for diarrhoea. The results of the first multicentre study in Italy to evaluate the potential clinical application of the GPP assay in the laboratory diagnosis of diarrhoea are reported here. METHODS: Faeces specimens (N=664) from hospitalized patients were tested with the GPP assay using a Luminex 200 instrument. All specimens were run using comparator methods following a routine algorithm: culture for bacteria, enzyme immunoassay and PCR for viruses, and microscopy for parasites. RESULTS: Of the samples tested with the GPP, 53.61% (356/664) gave positive results, as compared to 45.33% by routine testing. Of the positive specimens, 34.55% showed the presence of genomic DNA from multiple pathogens. The Luminex method showed an increase in the percentage of positivity of 8.28%. CONCLUSIONS: The GPP assay can be considered a helpful tool for the detection of gastrointestinal pathogens, with a hands-on time of 5h; it provides accurate data for the clinical management of hospitalized patients and for epidemiological surveillance.
OBJECTIVE:Gastroenteritis caused by a single pathogen or multiple pathogens remains a major diagnostic challenge for the laboratory. The treatment of diarrhoea is based on microbiological results. Diagnosis is achieved using different laboratory techniques that have variable sensitivity and specificity. xTAG GPP is a new multiplex PCR assay that simultaneously detects 15 different pathogens responsible for diarrhoea. The results of the first multicentre study in Italy to evaluate the potential clinical application of the GPP assay in the laboratory diagnosis of diarrhoea are reported here. METHODS: Faeces specimens (N=664) from hospitalized patients were tested with the GPP assay using a Luminex 200 instrument. All specimens were run using comparator methods following a routine algorithm: culture for bacteria, enzyme immunoassay and PCR for viruses, and microscopy for parasites. RESULTS: Of the samples tested with the GPP, 53.61% (356/664) gave positive results, as compared to 45.33% by routine testing. Of the positive specimens, 34.55% showed the presence of genomic DNA from multiple pathogens. The Luminex method showed an increase in the percentage of positivity of 8.28%. CONCLUSIONS: The GPP assay can be considered a helpful tool for the detection of gastrointestinal pathogens, with a hands-on time of 5h; it provides accurate data for the clinical management of hospitalized patients and for epidemiological surveillance.
Authors: Vu Thuy Duong; Voong Vinh Phat; Ha Thanh Tuyen; Tran Thi Ngoc Dung; Pham Duc Trung; Pham Van Minh; Le Thi Phuong Tu; James I Campbell; Hoang Le Phuc; Ton Thi Thanh Ha; Nguyen Minh Ngoc; Nguyen Thi Thanh Huong; Pham Thi Thanh Tam; Dang Thao Huong; Nguyen Van Xang; Nguyen Dong; Le Thi Phuong; Nguyen Van Hung; Bui Duc Phu; Tran My Phuc; Guy E Thwaites; Lu Lan Vi; Maia A Rabaa; Corinne N Thompson; Stephen Baker Journal: J Clin Microbiol Date: 2016-02-10 Impact factor: 5.948