F Morio1, S Valot2, A Laude3, G Desoubeaux4, N Argy5, C Nourrisson6, C Pomares7, M Machouart8, Y Le Govic9, F Dalle2, F Botterel10, N Bourgeois11, E Cateau12, M Leterrier13, F Jeddi3, M Gaboyard14, P Le Pape3. 1. Laboratoire de Parasitologie-Mycologie, Institut de Biologie, CHU Nantes, France. Electronic address: florent.morio@chu-nantes.fr. 2. Laboratoire de Parasitologie-Mycologie, CHU de Dijon, France. 3. Laboratoire de Parasitologie-Mycologie, Institut de Biologie, CHU Nantes, France. 4. Service de Parasitologie-Mycologie-Médecine Tropicale, CHU de Tours, CEPR Inserm U1100, équipe 3, Université François-Rabelais de Tours, France. 5. Laboratoire de Parasitologie-Mycologie, AP-HP, Hôpital Bichat, Paris, France. 6. Laboratoire de Parasitologie-Mycologie, CHU de Clermont-Ferrand, France. 7. Laboratoire de Parasitologie-Mycologie, INSERM, U1065, C3M, CHU de Nice, France. 8. Laboratoire de Parasitologie-Mycologie, CHU de Nancy, France. 9. Laboratoire de Parasitologie-Mycologie, CHU d'Angers, France. 10. Laboratoire de Parasitologie-Mycologie, AP-HP Hôpital Henri Mondor, Créteil, France. 11. Laboratoire de Parasitologie-Mycologie, CHU de Montpellier, France. 12. Laboratoire de Parasitologie-Mycologie, CHU de Poitiers, France. 13. Laboratoire de Microbiologie, CHD de la Roche-Sur-Yon, France. 14. Ademtech, Pessac, France.
Abstract
OBJECTIVES: Besides the potential to identify a wide variety of gastrointestinal parasites, microscopy remains the reference standard in clinical microbiology for amoeba species identification and, especially when coupled with adhesin detection, to discriminate the pathogenic Entamoeba histolytica from its sister but non-pathogenic species Entamoeba dispar/Entamoeba moshkovskii. However, this approach is time-consuming, requires a high-level of expertise that can be jeopardized considering the low prevalence of gastrointestinal parasites in non-endemic countries. Here, we evaluated the CE-IVD-marked multiplex PCR (ParaGENIE G-Amoeba, Ademtech) targeting E. histolytica and E. dispar/E. moshkovskii and Giardia intestinalis. METHODS: This evaluation was performed blindly on a reference panel of 172 clinical stool samples collected prospectively from 12 laboratories and analysed using a standardized protocol relying on microscopy (and adhesin detection by ELISA for the detection of E. histolytica) including G. intestinalis (n = 37), various amoeba species (n = 55) including E. dispar (n = 15), E. histolytica (n = 5), as well as 17 other gastrointestinal parasites (n = 80), and negative samples (n = 37). RESULTS: This new multiplex PCR assay offers fast and reliable results with appropriate sensitivity and specificity for the detection of G. intestinalis and E. dispar/E. moshkovskii from stools (89.7%/96.9% and 95%/100%, respectively). Detection rate and specificity were greatly improved by the PCR assay, highlighting several samples misidentified by microscopy, including false-negative and false-positive results for both E. dispar/E. moshkovskii and E. histolytica. CONCLUSION: Given the clinical relevance of amoeba species identification, microbiologists should be aware of the limitations of using an algorithm relying on microscopy coupled with adhesin detection by ELISA.
OBJECTIVES: Besides the potential to identify a wide variety of gastrointestinal parasites, microscopy remains the reference standard in clinical microbiology for amoeba species identification and, especially when coupled with adhesin detection, to discriminate the pathogenic Entamoeba histolytica from its sister but non-pathogenic species Entamoeba dispar/Entamoeba moshkovskii. However, this approach is time-consuming, requires a high-level of expertise that can be jeopardized considering the low prevalence of gastrointestinal parasites in non-endemic countries. Here, we evaluated the CE-IVD-marked multiplex PCR (ParaGENIE G-Amoeba, Ademtech) targeting E. histolytica and E. dispar/E. moshkovskii and Giardia intestinalis. METHODS: This evaluation was performed blindly on a reference panel of 172 clinical stool samples collected prospectively from 12 laboratories and analysed using a standardized protocol relying on microscopy (and adhesin detection by ELISA for the detection of E. histolytica) including G. intestinalis (n = 37), various amoeba species (n = 55) including E. dispar (n = 15), E. histolytica (n = 5), as well as 17 other gastrointestinal parasites (n = 80), and negative samples (n = 37). RESULTS: This new multiplex PCR assay offers fast and reliable results with appropriate sensitivity and specificity for the detection of G. intestinalis and E. dispar/E. moshkovskii from stools (89.7%/96.9% and 95%/100%, respectively). Detection rate and specificity were greatly improved by the PCR assay, highlighting several samples misidentified by microscopy, including false-negative and false-positive results for both E. dispar/E. moshkovskii and E. histolytica. CONCLUSION: Given the clinical relevance of amoeba species identification, microbiologists should be aware of the limitations of using an algorithm relying on microscopy coupled with adhesin detection by ELISA.
Authors: Silvia Paulos; José María Saugar; Aida de Lucio; Isabel Fuentes; María Mateo; David Carmena Journal: PLoS One Date: 2019-04-08 Impact factor: 3.240