Document Detail


Real-time PCR assay for rapid qualitative and quantitative detection of Entamoeba histolytica.
MedLine Citation:
PMID:  23195553     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Simple real-time PCR assay with one set of primer and probe for rapid, sensitive qualitative and quantitative detection of Entamoeba histolytica has been used. Consensus sequences were used to amplify a species-specific region of the 16S rRNA gene, and fluorescence resonance energy transfer hybridization probes were used for detection in a LightCycler platform (Roche). The anchor probe sequence was designed to be a perfect match for the 16S rRNA gene of Entamoeba species, while the acceptor probe sequence was designed for Entamoeba histolytica, which allowed differentiation. The performed characteristics of the real-time PCR assay were compared with ELISA antigen and microscopical detection from 77 samples of individuals with suspected clinical diagnosis of imported E. histolytica infection. Stool and liver abscess pus samples were examined with analytical sensitivity of 5 parasites per PCR reaction. The melting curve means Tms (standard deviation) in clinical isolates were 54°C. The real-time assay was 100% sensitive and specific for differentiation of Entamoeba histolytica, compared with conventional ELISA or microscopy. This real-time PCR assay with melting curve analysis is rapid, and specific for the detection and differentiation of Entamoeba histolytica. The suitability for routine use of this assay in clinical diagnostic laboratories is discussed.
Authors:
Erika Orosz; Katalin Perkátai; Beatrix Kapusinszky; Agnes Farkas; István Kucsera
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Acta microbiologica et immunologica Hungarica     Volume:  59     ISSN:  1217-8950     ISO Abbreviation:  Acta Microbiol Immunol Hung     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-11-30     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9434021     Medline TA:  Acta Microbiol Immunol Hung     Country:  Hungary    
Other Details:
Languages:  eng     Pagination:  451-60     Citation Subset:  IM    
Affiliation:
National Center for Epidemiology Department of Parasitology Gyáli út 2-6 H-1097 Budapest Hungary.
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