Improved diagnosis of gastrointestinal infections using a semi-automated multiplex real-time PCR for detection of enteropathogens

Introduction. The identification of enteropathogens is critical for the clinical management of patients with suspected gastrointestinal infection. The FLOW multiplex PCR system (FMPS) is a semi-automated platform (FLOW System, Roche) for multiplex real -time PCR analysis. Hypothesis/Gap Statement. F...

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Veröffentlicht in:Journal of medical microbiology 2021-01, Vol.70 (9), Article 001367
Hauptverfasser: Fidalgo, Berta, Rubio, Elisa, Pastor, Victor, Parera, Marta, Balleste-Delpierre, Clara, Fernandez, Mariana Jose, Chasco, Genoveva Cuesta, Vergara, Andrea, Zboromyrska, Yuliya, Aylagas, Cristian, Salvador, Pilar, Fernandez, Adan, Valls, M. Eugenia, Martinez, Miriam Jose Alvarez, Mira, Aurea, Marcos, Maria Angeles, Vila, Jordi, Martinez, Miguel J., Casals-Pascual, Climent
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container_issue 9
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container_title Journal of medical microbiology
container_volume 70
creator Fidalgo, Berta
Rubio, Elisa
Pastor, Victor
Parera, Marta
Balleste-Delpierre, Clara
Fernandez, Mariana Jose
Chasco, Genoveva Cuesta
Vergara, Andrea
Zboromyrska, Yuliya
Aylagas, Cristian
Salvador, Pilar
Fernandez, Adan
Valls, M. Eugenia
Martinez, Miriam Jose Alvarez
Mira, Aurea
Marcos, Maria Angeles
Vila, Jordi
Martinez, Miguel J.
Casals-Pascual, Climent
description Introduction. The identification of enteropathogens is critical for the clinical management of patients with suspected gastrointestinal infection. The FLOW multiplex PCR system (FMPS) is a semi-automated platform (FLOW System, Roche) for multiplex real -time PCR analysis. Hypothesis/Gap Statement. FMPS has greater sensitivity for the detection of enteric pathogens than standard methods such as culture, biochemical identification, immunochromatography or microscopic examination. Aim.The diagnostic performance of the FMPS was evaluated and compared to that of traditional microbiological procedures. Methodology. A total of 10 659 samples were collected and analysed over a period of 7 years. From 2013 to 2018 (every July to September), samples were processed using standard microbiological culture methods. In 2019, the FMPS was implemented using real -time PCR to detect the following enteropathogens: Shigella spp., Salmonella spp., Campylobacter spp., Giardia intestinalis, Entamoeba histolytica, Blastocystis hominis, Cryptosporidum spp., Dientamoeba fragilis, adenovirus, norovirus and rotavirus. Standard microbiological culture methods (2013-2018) included stool culture, microscopy and immunochromatography. Results. A total of 1078 stool samples were analysed prospectively using the FMPS from July to September (2019): bacterial, parasitic and viral pathogens were identified in 15.3, 9.71 and 5.29 % of cases, respectively. During the same period of 6 years (2013-2018), the proportion of positive identifications using standard microbiological methods from 2013 to 2018 was significantly lower. A major significant recovery improvement was observed for all bacteria species tested: Shigella spp./enteroinvasive Escherichia coli (EIEC) (P
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Eugenia ; Martinez, Miriam Jose Alvarez ; Mira, Aurea ; Marcos, Maria Angeles ; Vila, Jordi ; Martinez, Miguel J. ; Casals-Pascual, Climent</creator><creatorcontrib>Fidalgo, Berta ; Rubio, Elisa ; Pastor, Victor ; Parera, Marta ; Balleste-Delpierre, Clara ; Fernandez, Mariana Jose ; Chasco, Genoveva Cuesta ; Vergara, Andrea ; Zboromyrska, Yuliya ; Aylagas, Cristian ; Salvador, Pilar ; Fernandez, Adan ; Valls, M. Eugenia ; Martinez, Miriam Jose Alvarez ; Mira, Aurea ; Marcos, Maria Angeles ; Vila, Jordi ; Martinez, Miguel J. ; Casals-Pascual, Climent</creatorcontrib><description>Introduction. The identification of enteropathogens is critical for the clinical management of patients with suspected gastrointestinal infection. The FLOW multiplex PCR system (FMPS) is a semi-automated platform (FLOW System, Roche) for multiplex real -time PCR analysis. Hypothesis/Gap Statement. FMPS has greater sensitivity for the detection of enteric pathogens than standard methods such as culture, biochemical identification, immunochromatography or microscopic examination. Aim.The diagnostic performance of the FMPS was evaluated and compared to that of traditional microbiological procedures. Methodology. A total of 10 659 samples were collected and analysed over a period of 7 years. From 2013 to 2018 (every July to September), samples were processed using standard microbiological culture methods. In 2019, the FMPS was implemented using real -time PCR to detect the following enteropathogens: Shigella spp., Salmonella spp., Campylobacter spp., Giardia intestinalis, Entamoeba histolytica, Blastocystis hominis, Cryptosporidum spp., Dientamoeba fragilis, adenovirus, norovirus and rotavirus. Standard microbiological culture methods (2013-2018) included stool culture, microscopy and immunochromatography. Results. A total of 1078 stool samples were analysed prospectively using the FMPS from July to September (2019): bacterial, parasitic and viral pathogens were identified in 15.3, 9.71 and 5.29 % of cases, respectively. During the same period of 6 years (2013-2018), the proportion of positive identifications using standard microbiological methods from 2013 to 2018 was significantly lower. A major significant recovery improvement was observed for all bacteria species tested: Shigella spp./enteroinvasive Escherichia coli (EIEC) (P &lt;0.05), Salmonella spp. (P &lt;0.05) and Campylobacter spp. (P &lt;0.05). Marked differences were also observed for the parasites G. intestinalis, Cryptosporidium spp. and D. fragilis. Conclusion. These results support the value of multiplex real -time PCR analysis for the detection of enteric pathogens in laboratory diagnosis with outstanding performance in identifying labile microorganisms. The identification of unsuspected microorganisms for less specific clinical presentations may also impact on clinical practice and help optimize patient management.</description><identifier>ISSN: 0022-2615</identifier><identifier>EISSN: 1473-5644</identifier><identifier>DOI: 10.1099/jmm.0.001367</identifier><identifier>PMID: 34516365</identifier><language>eng</language><publisher>LONDON: Microbiology Soc</publisher><subject>Life Sciences &amp; Biomedicine ; Microbiology ; Science &amp; Technology</subject><ispartof>Journal of medical microbiology, 2021-01, Vol.70 (9), Article 001367</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>true</woscitedreferencessubscribed><woscitedreferencescount>6</woscitedreferencescount><woscitedreferencesoriginalsourcerecordid>wos000705457900015</woscitedreferencesoriginalsourcerecordid><citedby>FETCH-LOGICAL-c268t-f720424c4a872fade99ba0cea123d8d9da42c90080051eb3f7119715825549813</citedby><cites>FETCH-LOGICAL-c268t-f720424c4a872fade99ba0cea123d8d9da42c90080051eb3f7119715825549813</cites><orcidid>0000-0002-0682-1075 ; 0000-0002-0867-8954 ; 0000-0002-1685-7127 ; 0000-0001-8589-2519 ; 0000-0003-2716-4729 ; 0000-0002-2556-8837 ; 0000-0002-9031-0787 ; 0000-0003-2527-3956</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,781,785,3747,27929,27930,39263</link.rule.ids></links><search><creatorcontrib>Fidalgo, Berta</creatorcontrib><creatorcontrib>Rubio, Elisa</creatorcontrib><creatorcontrib>Pastor, Victor</creatorcontrib><creatorcontrib>Parera, Marta</creatorcontrib><creatorcontrib>Balleste-Delpierre, Clara</creatorcontrib><creatorcontrib>Fernandez, Mariana Jose</creatorcontrib><creatorcontrib>Chasco, Genoveva Cuesta</creatorcontrib><creatorcontrib>Vergara, Andrea</creatorcontrib><creatorcontrib>Zboromyrska, Yuliya</creatorcontrib><creatorcontrib>Aylagas, Cristian</creatorcontrib><creatorcontrib>Salvador, Pilar</creatorcontrib><creatorcontrib>Fernandez, Adan</creatorcontrib><creatorcontrib>Valls, M. Eugenia</creatorcontrib><creatorcontrib>Martinez, Miriam Jose Alvarez</creatorcontrib><creatorcontrib>Mira, Aurea</creatorcontrib><creatorcontrib>Marcos, Maria Angeles</creatorcontrib><creatorcontrib>Vila, Jordi</creatorcontrib><creatorcontrib>Martinez, Miguel J.</creatorcontrib><creatorcontrib>Casals-Pascual, Climent</creatorcontrib><title>Improved diagnosis of gastrointestinal infections using a semi-automated multiplex real-time PCR for detection of enteropathogens</title><title>Journal of medical microbiology</title><addtitle>J MED MICROBIOL</addtitle><description>Introduction. The identification of enteropathogens is critical for the clinical management of patients with suspected gastrointestinal infection. The FLOW multiplex PCR system (FMPS) is a semi-automated platform (FLOW System, Roche) for multiplex real -time PCR analysis. Hypothesis/Gap Statement. FMPS has greater sensitivity for the detection of enteric pathogens than standard methods such as culture, biochemical identification, immunochromatography or microscopic examination. Aim.The diagnostic performance of the FMPS was evaluated and compared to that of traditional microbiological procedures. Methodology. A total of 10 659 samples were collected and analysed over a period of 7 years. From 2013 to 2018 (every July to September), samples were processed using standard microbiological culture methods. In 2019, the FMPS was implemented using real -time PCR to detect the following enteropathogens: Shigella spp., Salmonella spp., Campylobacter spp., Giardia intestinalis, Entamoeba histolytica, Blastocystis hominis, Cryptosporidum spp., Dientamoeba fragilis, adenovirus, norovirus and rotavirus. Standard microbiological culture methods (2013-2018) included stool culture, microscopy and immunochromatography. Results. A total of 1078 stool samples were analysed prospectively using the FMPS from July to September (2019): bacterial, parasitic and viral pathogens were identified in 15.3, 9.71 and 5.29 % of cases, respectively. During the same period of 6 years (2013-2018), the proportion of positive identifications using standard microbiological methods from 2013 to 2018 was significantly lower. A major significant recovery improvement was observed for all bacteria species tested: Shigella spp./enteroinvasive Escherichia coli (EIEC) (P &lt;0.05), Salmonella spp. (P &lt;0.05) and Campylobacter spp. (P &lt;0.05). Marked differences were also observed for the parasites G. intestinalis, Cryptosporidium spp. and D. fragilis. Conclusion. These results support the value of multiplex real -time PCR analysis for the detection of enteric pathogens in laboratory diagnosis with outstanding performance in identifying labile microorganisms. 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Eugenia</au><au>Martinez, Miriam Jose Alvarez</au><au>Mira, Aurea</au><au>Marcos, Maria Angeles</au><au>Vila, Jordi</au><au>Martinez, Miguel J.</au><au>Casals-Pascual, Climent</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Improved diagnosis of gastrointestinal infections using a semi-automated multiplex real-time PCR for detection of enteropathogens</atitle><jtitle>Journal of medical microbiology</jtitle><stitle>J MED MICROBIOL</stitle><date>2021-01-01</date><risdate>2021</risdate><volume>70</volume><issue>9</issue><artnum>001367</artnum><issn>0022-2615</issn><eissn>1473-5644</eissn><abstract>Introduction. The identification of enteropathogens is critical for the clinical management of patients with suspected gastrointestinal infection. The FLOW multiplex PCR system (FMPS) is a semi-automated platform (FLOW System, Roche) for multiplex real -time PCR analysis. Hypothesis/Gap Statement. FMPS has greater sensitivity for the detection of enteric pathogens than standard methods such as culture, biochemical identification, immunochromatography or microscopic examination. Aim.The diagnostic performance of the FMPS was evaluated and compared to that of traditional microbiological procedures. Methodology. A total of 10 659 samples were collected and analysed over a period of 7 years. From 2013 to 2018 (every July to September), samples were processed using standard microbiological culture methods. In 2019, the FMPS was implemented using real -time PCR to detect the following enteropathogens: Shigella spp., Salmonella spp., Campylobacter spp., Giardia intestinalis, Entamoeba histolytica, Blastocystis hominis, Cryptosporidum spp., Dientamoeba fragilis, adenovirus, norovirus and rotavirus. Standard microbiological culture methods (2013-2018) included stool culture, microscopy and immunochromatography. Results. A total of 1078 stool samples were analysed prospectively using the FMPS from July to September (2019): bacterial, parasitic and viral pathogens were identified in 15.3, 9.71 and 5.29 % of cases, respectively. During the same period of 6 years (2013-2018), the proportion of positive identifications using standard microbiological methods from 2013 to 2018 was significantly lower. A major significant recovery improvement was observed for all bacteria species tested: Shigella spp./enteroinvasive Escherichia coli (EIEC) (P &lt;0.05), Salmonella spp. (P &lt;0.05) and Campylobacter spp. (P &lt;0.05). Marked differences were also observed for the parasites G. intestinalis, Cryptosporidium spp. and D. fragilis. Conclusion. These results support the value of multiplex real -time PCR analysis for the detection of enteric pathogens in laboratory diagnosis with outstanding performance in identifying labile microorganisms. 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title Improved diagnosis of gastrointestinal infections using a semi-automated multiplex real-time PCR for detection of enteropathogens
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