№ files_lp_4_process_1_42884
Supplementary laboratory protocol presenting primer and probe sequences, reaction compositions, thermocycling parameters, plasmid constructs, and validation results for a real-time PCR panel targeting multiple intestinal protozoa in stool samples.
Year: Not specified
Region / City: Basel, Switzerland
Field: Parasitology
Document Type: Supplementary scientific protocol and validation data
Institution: Swiss Tropical and Public Health Institute (SwissTPH)
Department: National Reference Centre for Parasitology
Methodology: Real-time PCR using TaqMan probe assays
Assay Format: Six duplex real-time PCR reactions and one simplex reaction
Target Organisms: Entamoeba dispar, Entamoeba histolytica, Cryptosporidium spp., Cryptosporidium hominis/parvum, Microsporidia (Encephalitozoon spp., Enterocytozoon bieneusi), Entamoeba moshkovskii, Entamoeba polecki, Dientamoeba fragilis, Giardia lamblia, Cyclospora cayetanensis, Cystoisospora belli, Blastocystis sp.
Gene Target: 18S rRNA gene
Sample Type: Human stool samples
Validation Material: Plasmids with sequence inserts and clinical samples
Analytical Sensitivity: 1–10 plasmids/µl depending on assay
Diagnostic Sensitivity: Reported up to 100% for validated assays
Specificity: 100% for tested organisms during validation
Laboratory Method: Quantitative real-time polymerase chain reaction (qPCR)
Thermocycling Profile: 50 °C for 2 min, 95 °C for 10 min, followed by 40 cycles of 95 °C for 15 s and 58 °C for 1 min
DNA Extraction Material: Approximately 1 g stool aliquots stored at −80 °C prior to molecular analysis
Reagents: GeneExpression Mastermix (Thermo Fisher Scientific), primers and TaqMan probes
Price: 8 / 10 USD
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