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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.
PLoS One. 2019; 14(4):e0215068.Plos

Abstract

BACKGROUND

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.

METHODS

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).

PRINCIPAL FINDINGS

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.

CONCLUSIONS

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.

Links

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Authors+Show Affiliations

Paulos S
Synlab Group, Microbiology Service, Quirón Madrid University Hospital, Pozuelo de Alarcón, Madrid, Spain.
Saugar JM
Parasitology Reference and Research Laboratory, National Centre for Microbiology, Carlos III Health Institute, Majadahonda, Madrid, Spain.
de Lucio A
Parasitology Reference and Research Laboratory, National Centre for Microbiology, Carlos III Health Institute, Majadahonda, Madrid, Spain.
Fuentes I
Parasitology Reference and Research Laboratory, National Centre for Microbiology, Carlos III Health Institute, Majadahonda, Madrid, Spain.
Mateo M
European University of Madrid, Villaviciosa de Odón, Madrid, Spain.
Carmena D
Parasitology Reference and Research Laboratory, National Centre for Microbiology, Carlos III Health Institute, Majadahonda, Madrid, Spain.

MeSH

Biological AssayCryptosporidiosisCryptosporidium parvumDiagnostic Tests, RoutineDiarrheaEntamoeba histolyticaEntamoebiasisFecesGiardia lambliaGiardiasisHumansMultiplex Polymerase Chain Reaction

Pub Type(s)

Comparative Study
Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

30958837

Citation

Paulos, Silvia, et al. "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." PloS One, vol. 14, no. 4, 2019, pp. e0215068.
Paulos S, Saugar JM, de Lucio A, et al. 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. PLoS One. 2019;14(4):e0215068.
Paulos, S., Saugar, J. M., de Lucio, A., Fuentes, I., Mateo, M., & Carmena, D. (2019). 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. PloS One, 14(4), e0215068. https://doi.org/10.1371/journal.pone.0215068
Paulos S, et al. 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. PLoS One. 2019;14(4):e0215068. PubMed PMID: 30958837.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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. AU - Paulos,Silvia, AU - Saugar,José María, AU - de Lucio,Aida, AU - Fuentes,Isabel, AU - Mateo,María, AU - Carmena,David, Y1 - 2019/04/08/ PY - 2018/11/02/received PY - 2019/03/26/accepted PY - 2019/4/9/entrez PY - 2019/4/9/pubmed PY - 2019/12/28/medline SP - e0215068 EP - e0215068 JF - PloS one JO - PLoS One VL - 14 IS - 4 N2 - BACKGROUND: 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. METHODS: 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). PRINCIPAL FINDINGS: 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. CONCLUSIONS: 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. SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/30958837/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_ DB - PRIME DP - Unbound Medicine ER -