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Bacteria Plus kit. First, each lot of primer/probe was combined with each lot of Master Mix
and all combinations were tested with a single DNA Bacteria kit lot(see Table 6) on a single
Rotor-Gene Q instrument.
Table 6. Evaluation of
mericon
E.coliO157 Screen Plus and
mericon
E.coliSTEC O-typeLot-
to-lot Variability
Primer/Probes Lot 1
Lot 2
Lot 3
Lot 1
Lot 2
Lot 3
Lot 1
Lot 2
Lot 3
Master Mix
Lot 1
Lot 1
Lot 1
Lot 2
Lot 2
Lot 2
Lot 3
Lot 3
Lot 3
DNA Bacteria Lot 1
Lot 1
Lot 1
Lot 1
Lot 1
Lot 1
Lot 1
Lot 1
Lot 1
Second, each of 3 lots of DNA Bacteria kits was tested with a single lot of
mericon
E.coliO157Screen Plus or
mericon
E.coliSTEC O-type PCR kits primer/probe and
master mix (see Table 7). At the same time, three Rotor-Gene Q instruments were tested
side-by-side with the same kit lots.
Table 7. Evaluation of DNA Bacteria Plus and QIAsymphony Lot-to-lot Variability and
Instrument Variability
Bacterial strains used for the
mericon
E. coliO157 Screen Plus lot to lot variation studies
wereE. coli strains O157:H7 (RM-HYG374), O157(RM-HYG381), O26:H (RM-HYG375), E.
coli O111:H- (RM-YG376)and
Citrobacter freundii
(ifp culture collection)
Bacterial strains used for the
mericon
E.coliSTEC O-type lot to lot variation studies were
E. coli strainsO157:H7 (RM-HYG374), O157(RM-HYG381),E.coli O26:H- (RM-HYG375),
E. coli O111:H- (RM-YG376),E.coli (O145 RM-HYG390)and
Citrobacter freundii
.
Stability testing: Shelf life
Product stability data was submitted with the QIAGEN
mericon
Salmonella study (July
2012). For the purposes of this study, shelf life stability studies are being submitted for the
E. coli
Screen Plusand
E. coli
STEC O-typeprimer probe sets, since these are the only
components of the workflow protocols that vary between the
mericon
protocols. The
Salmonella
summary data can be found in Appendix 2 of the current report. Bacterial
strains used were
Salmonellaenterica
subsp
enterica
serovar Abony
(NCTC
6017)
,
Salmonella enterica
subsp
enterica
serovar Enteriditis, (Sm24/Rif12/Ssq),
Citrobacter
freundii, (ifp culture collection).
mericon
E.coliO157 Screen Plus and
mericon
E.coliSTEC O-typepathogen kit stability
studies
Primer/Probes
Lot 1
Lot 1
Lot 1
Lot 1
Lot 1
Master Mix
Lot 1
Lot 1
Lot 1
Lot 1
Lot 1
DNA Bacteria
Lot 1
Lot 1
Lot 1
Lot 2
Lot 3
Rotor-Gene Q
RGQ #1 RGQ #2 RGQ #3 RGQ #1 RGQ #1
OMAMAN-36 E/ AOAC PTM Report
ERP Use Only
January 2017
AOAC Research Institute
Expert Review Panel Use Only