Comparative performance evaluation of four commercial multiplex real-time PCR assays for the detection of the diarrhoea-causing protozoa Cryptosporidium hominis/parvum, Giardia duodenalis and Entamoeba histolytica

• Published in: PloS one Vol. 14; no. 4; p. e0215068
• Main Authors: Paulos, Silvia, Saugar, José María, de Lucio, Aida, Fuentes, Isabel, Mateo, María, Carmena, David
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 08.04.2019; Public Library of Science (PLoS)
• Subjects: Algorithms; Biological Assay; Biology and Life Sciences; Cryptosporidiosis - diagnosis; Cryptosporidiosis - parasitology; Cryptosporidium; Cryptosporidium hominis; Cryptosporidium parvum; Cryptosporidium parvum - genetics; Cryptosporidium parvum - isolation & purification; Deoxyribonucleic acid; Diagnosis; Diagnostic systems; Diagnostic Tests, Routine - methods; Diarrhea; Diarrhea - diagnosis; Diarrhea - epidemiology; Diarrhea - parasitology; DNA; Entamoeba histolytica; Entamoeba histolytica - genetics; Entamoeba histolytica - isolation & purification; Entamoebiasis - diagnosis; Entamoebiasis - parasitology; Feces - parasitology; Frontotemporal dementia; Gastroenteritis; Giardia; Giardia duodenalis; Giardia lamblia - genetics; Giardia lamblia - isolation & purification; Giardiasis - diagnosis; Giardiasis - parasitology; Humans; Laboratories; Medical diagnosis; Medicine and Health Sciences; Multiplex Polymerase Chain Reaction - methods; Multiplexing; Parasites; Parasitic diseases; Pathogenic microorganisms; Pathogens; Performance evaluation; Polymerase chain reaction; Protozoa; Real time; Research and Analysis Methods; Sensitivity; Test procedures; Workflow; Workflow software; United States > US; Europe; Spain
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0215068

Multiplex molecular panels are relentlessly replacing conventional methods for the detection of enteric pathogens from stool samples in clinical and research laboratories. Here we evaluated four commercial multiplex real-time PCR assays for the detection of Cryptosporidium hominis/parvum, Giardia duodenalis and Entamoeba histolytica. The diagnostic performance of the Gastroenteritis/Parasite Panel I (Diagenode), the RIDAGENE Parasitic Stool Panel (R-Biopharm), the Allplex Gastrointestinal Parasite Panel 4 (Seegene) and the FTD Stool Parasites (Fast Track) real-time PCR methods was assessed against a reference panel of 126 well-characterized DNA samples including Cryptosporidium hominis (n = 29), Cryptosporidium parvum (n = 3), Giardia duodenalis (n = 47), Entamoeba histolytica (n = 3), other parasite species (n = 20), and apparently healthy subjects (n = 24). Obtained diagnostic sensitivities ranged from 53-88% for Cryptosporidium hominis/parvum, and from 68-100% for G. duodenalis. The R-Biopharm method achieved the best performance for the detection of Cryptosporidium hominis/parvum both in terms of diagnostic sensitivity (87.5%) and detection limit (a 100-fold increase compared to other tests). The Fast Track method was particularly suited for the detection of G. duodenalis, achieving a 100% sensitivity and a detection limit at least 10-fold superior. Detection of E. histolytica was similarly achieved by all compared methods except Diagenode. Diagnostic performance varied largely depending on the method used and the targeted pathogen species. Factors including test sensitivity/specificity, cost, patient population surveyed, laboratory workflow, and diagnostic algorithm should be carefully considered when choosing the most appropriate multiplex PCR platform.