1055 / 1195
MOZ,OLA
I]T
AI.,.: JOURNAI-
O}.
AOAC
INI.EIìNÂI'IONAI-
VOL. 97'
NO.
3,
20I1
829
FOOD
BIOLOGICAL
CONTAMINANTS
Evaluation
of
the
AI{SR.
for
Sulmonellø
Assay
for
Identification
of Sulmonella
spp.
from
Colony Picks
from
Selective/Differential
Agar
Media: First Action
2013.14
jVl;rnn
Mozor,;r,
M;\Xtì\{lLtAN
Bot't;rrrn, C..rnolyx J.rcaulcs,
Plul
NORrox, Osc;rn
CÁBÄLl,ERol
NIcol-E
ENSLtx'
PR¡¡;r'll,r
B¡srvrs,
and JDxNltrER
Rlcti
Neogen Corp., 620
l.,esher
Pl,
L.ansing.
MI
48912
Collaborators:
E.S.
Adanls.
H.
Alnughal,mishi,
J.B.
Barrett,
J. Benz.inger,
M.E.
Ilen'ang,
M.
ISoyle, J.
Cannon. D.
C'lark,
A.
Copeland,
M.
Corebello, D.E.
Cosby,
N.A.
Cox,
N.
Cuthbert,
ll.
Danrntann, J.
Dyszel, E.
Feldpausch, J. Flannery.
C.
Flores.
J.C. Frye,
Iì.
Fuller,
V.
Gill.
1...M.
I-liott,
Il.
I-lorvard,
M.
Hudgens,
C.R.
Jackson,
W. Jones, S.W.
Knapp.
L.
Kuepler,
Iì
Kulkarni'
B. Kupski, C.
Pidgeon,
A.
Quenneville,
L.L.
Rigsby,
N.
Rogrnan,
E.
Sai,
Â.
Scollon, lvl.
Sisemore, J.
Stepnitz.
J.
Van
llrunt,
M.
Vross, D.
\\hltman,
Iì.
Wang.
G.
Whitbeck,
S.
York,
l'.
Zhang
A
collaborative study
was
conducted to
evaluate
performance of
the
ANSR@
for
Sa/monel/a assay
for
identification of Salmonella spp. from colony picks
lakcn frnm selpcfive/r{ifferential
aoar media- The
ANSR Sa/monel/a
assay is
an
isothermal nucleic
acid amplification
test
based on
the nicking
enzyme
amplification reaction chemistry.
The
test can
be
completed in less than
40
min including sample
preparation.
A
total
of
18
laboratories representing
industry, government,
academic, and
commercial
testing
laboratories participated
in the
study.
Each
collaborator tested
up
to
84
samples, comprised
of
colony picks of
six
Sa/¡nonel/a
spp.
and
six
non-
sa lm
onel lae
taken from
s
ix selective/differential
agar media as
well
as
tryptic
soy
agar, A
total of
12t41
analyses were performed,
1416
oî
which
gave
the correct identification,
for
overall accuracy
of
98.3%.
For
identification
of
Salmonella
spp.,
755
of
756
tests
(99.9%)
produced the correct result.
For
identification
of non-salmonellae
as
such,
661 of
685
assays
(96.5%)
produced the correct result,
Of
the
lS
laboratories,
15
produced
data sets
with
99-100%
accuracy.
The
majoriÇ
of false-positive results
were
clustered in three laboratories; analysis of
raw data
suggests procedural
difficulties
in
at least
two
cases,
which
may
explain the atypical
data
from these
collaborators.
The
ANSR Sa/monel/a assay
can
be
used as a
rapid, accurate adjunct
or
alternative to
biochemical
testing for
identification of presumptive
Salmonell
a
spp. isolates.
lìeceived Decenrber
I
8.
201 3.
The nrethori
u,a.s
approve
tl
b),
the ]:\pe
rt llevie\\'
Panel
lor
lrood
lìiological
Cont¿ìminants
.1s
l:irst
Action.
'l-he
Iìrpert
lìevierl'
Panel
1'or
Food
Biological
Contaminants
invites
nethod
users
to
provide
leedback on
the i'-irst
Action
methods
Ireedback
fronr
nretliod
users
rvill
help
verifl,
that
the
nlethods
are
fìt for
purpose
and
are
critical
to gaining global recognition
and
¿ìcceptance
of
tlìe
lrethods.
Cìournlsnts
call
be sent
directly
to
thc
corresponding author or
nlethodfeedback@aoac.org.Corresponding author's
e-rrail.
nlnrozola@neogen.comAn
appendix is available on
the
J.
¿1O,4C
lnt.
rvebsite, http://aoac.
publ
isher.
i
ngentâcon
nect.
com/contenVaoac/j
aoac
DOt:
10.5740/jaoacint. ì4-004
fdentifìcatiort
of
presumplive Salmonella
colonies
from
I
selective/differcntial agar media
as
Sal¡nonella
spp.
has
Ihistorically
been achiev"ed
using
a
variety of biochemical
and
serological
procedures.
In
the
case
of
f-ood
and
environmental
sanrple analysis, these
pt'ocedures
are
specifìed
in
reference
methods
such
as
those in the tJ.S. F'ood and
DrugAdministration's
BacÍeriological
Analytical
l\.lanttal
(BAMI
l)
and
the
I.J.S,
Department ofAgriculture's,41r
crobi
ol
og,
La
boraÍ
orv
Gui
de
book
(MLC;2),
These
trethods
include conventional biochemical
tests,
miniaturized
biochemical
test
devices, automated biochemical
identification platforms,
and
serological agglutination
tests
using
Salmonella-specific
antisera.
The
biochemical
ìdentification
procedures,
although
accurale
and reliable. generally
reqLrire
6-24
h
to
obtain
results.
The
serological procedures
may
be
rapid, but often
requile
subculture to
enhance
antigen expression.
especially
in thc
case
ofl'lagellar
(fl)
antigen
typing.
As
an adiunct
or
altemative
to
biochemical and
serological
procedures.
nucleic
acid-based identification rnethods
hold
pronrise
fbr
providing
timely
and accurate results.
'fhis
has been
acknorvledged,
flor
exarnple. by relerence in both
BAM
and
MLG
to
use
of
nucleic
acid-based rÌìethods
tbr
identificatiot't
of
I'isteria
nronocytogenes
(3,4).
The
ANSR@
Salnonellu
assa)/
was
originall¡,
developed fbr
rapid
screening
of
enriched
food and
environmental
samples.
The
assay
is
an
isothennal nucleic acid amplification
procedttre,
based
on
the nicking
enzyme
arnplification
reaction (NEAR)
technology (5). The
ANSIì
rnethod has been
cvaluated
in
thrce
AO
AC
P
e
r!'or
tn
a
nc
e'l
þ
s
r
e
d
14
erl'ofNa
1P'l'
M
;
val
id
ati
on
stucl
ies.
leading to
ceftilìcation
as
P'lM
061203.
rvith claims for
a
varictr
ol
I'ood and
environmental
sample types
(6-7),
In
these
str¡dies.
the
ANSIì
method
exhibited
sensitit'it--v
comparable
to
that
of'
the
BAM
and
MLG
rel'elence
culture
methocls
by
probabiìiry'
ol-<ietection
stalistical
analYsis.
as
u'ell
a-s
>99%o
inclusivity
and
1007o
exclusivity in
testing
oftarget
ancl
nontatget
bacteril.
'lhis
method perfolmance, coupled
rvith the simplicity
and
rapidity
o1'the assay (less
than
40
min),
suggested
th¿rt
the
rnethod
could
also serve
as
a
usefr¡l
tool
lbr
identilìcation ofpresunrptive
Salntonella spp.
isolates
liom
selective/diffelential agal plating
media.
A
precollaborative study
has
been conrpleted
in
rvhich
colorries
of
113
Salntonellct
spp.
strains and 37
non-Solntonella
strains
rvere
picked
floni
trvptic
soy
agar
('l'SA)
and
six selective/
difl'erential
agar
media
[-lektoen
enteric
agar (FIF.),
x¡'lose
lysine