AOAC ISPAM Meeting eBook, March 17 2015

Virus : detection method?

CEN/TC275/WG6/TAG 4 ISO/TS 15216

OPTIMIZED FOR ROUTINE ANALYSIS

ROBUST

FAST

SENSITIVE % AMPLIFICATION EFFICACY STANDARDIZED AMPLIFICATION S A MULTIPLEXING M 95

SPECIFIC

MULTI- PLATEFORMS

MULTI-MATRICES

EXTERNAL CONTROL MENGOVIRUS

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Virus : detection method?

CEN/TC275/WG6/TAG4 ISO/TS 15216

VALIDATED PROCEDURE DIAGRAM

DETECTION

VIRUS PRETREATMENT

MENGOVIRUS ADDED

EXTRACTION RNA

HARD SURFA CES SALADS VEGGIES FRUITS HERBS SPICES

100 cm 2

Nucleic acids extraction on 1mL concentrated virus, lysis with guanidine isothiocyanate & silica binding. Elution RNA in 100 µl (= MiniMag equipment + Nuclisens reagents bioMérieux)

SWABBING FOLLOWED BY ELUTION

GLYCINE BUFFER ELUTION PRECIPITATION PEG / NaCl CLARIFICATION CHLOROFORM BUTANOL DIGESTIVE TISSUES ELUTION PROTEINASE K INCUBATION 37 ° THEN 60 ° CENTRIFUGATION CONCENTRATION / FILTRATION TANGENTIAL CASSETTES, PRECIPITATION PEG CLARIFICATION CHLOROFORM BUTANOL

Mengovirus DETECTION & targets

25g

2g

BIVALVE MOLLUSKS

RESULTS INTERPRET ATION

1L

LIQUID SAMPLES

12