TY - JOUR
T1 - PCR revisited
T2 - A case for revalidation of PCR assays for microorganisms using identification of Campylobacter species as an exemplar
AU - On, S.L.W.
AU - Brandt, S.M.
AU - Cornelius, A.J.
AU - Fusco, V.
AU - Quero, G.M.
AU - Mackiw, E.
AU - Houf, K.
AU - Bilbao, A.
AU - Diaz, A.I.
AU - Benejat, L.
AU - Megraud, F.
AU - Collins-Emerson, J.
AU - French, N.P.
AU - Gotcheva, V.
AU - Angelov, A.
AU - Alakomi, Hanna-Leena
AU - Saarela, M.
AU - Paulin, S.M.
PY - 2013
Y1 - 2013
N2 - We re-examined the sensitivity and specificity of 31 PCR assays
(including four commercially available and developed in-house methods)
for the identification of Campylobacter species, with particular reference to taxa described since 2004, which are closely related to C. jejuni and C. coli,
the pathogenic species of most interest. Each of the assays was used by
at least one of the participating nine laboratories in eight countries.
The sensitivity and specificity of these PCR assays examined varied
considerably and ranged from 100% to 0% for sensitivity and 100% to 55%
for specificity. None of the three assays examined for C. lari were successful in detecting all strains of this species, possibly reflecting its complex taxonomy. A number of assays for C. jejuni, C. coli, and a subgroup of enteropathogenic campylobacters, were found to yield false positive results for Campylobacter species described since PCR tests were reported, including C. cuniculorum, C. subantarcticus, C. peloridis and C. volucris.
Our study supports the need for attention to detail in initial PCR
assay design and evaluation, and also for on-going revalidation of
laboratory assays to ensure that diagnoses are correct. Recommendations
to guide the revalidation process are presented.
AB - We re-examined the sensitivity and specificity of 31 PCR assays
(including four commercially available and developed in-house methods)
for the identification of Campylobacter species, with particular reference to taxa described since 2004, which are closely related to C. jejuni and C. coli,
the pathogenic species of most interest. Each of the assays was used by
at least one of the participating nine laboratories in eight countries.
The sensitivity and specificity of these PCR assays examined varied
considerably and ranged from 100% to 0% for sensitivity and 100% to 55%
for specificity. None of the three assays examined for C. lari were successful in detecting all strains of this species, possibly reflecting its complex taxonomy. A number of assays for C. jejuni, C. coli, and a subgroup of enteropathogenic campylobacters, were found to yield false positive results for Campylobacter species described since PCR tests were reported, including C. cuniculorum, C. subantarcticus, C. peloridis and C. volucris.
Our study supports the need for attention to detail in initial PCR
assay design and evaluation, and also for on-going revalidation of
laboratory assays to ensure that diagnoses are correct. Recommendations
to guide the revalidation process are presented.
KW - campylobacter coli
KW - campylobacter jejuni
KW - identification
KW - PCR
KW - revalidation
U2 - 10.3920/QAS2012.0158
DO - 10.3920/QAS2012.0158
M3 - Article
SN - 1757-8361
VL - 5
SP - 49
EP - 62
JO - Quality Assurance and Safety of Crops and Foods
JF - Quality Assurance and Safety of Crops and Foods
ER -