PCR revisited: A case for revalidation of PCR assays for microorganisms using identification of Campylobacter species as an exemplar

S.L.W. On (Corresponding Author), S.M. Brandt, A.J. Cornelius, V. Fusco, G.M. Quero, E. Mackiw, K. Houf, A. Bilbao, A.I. Diaz, L. Benejat, F. Megraud, J. Collins-Emerson, N.P. French, V. Gotcheva, A. Angelov, Hanna-Leena Alakomi, M. Saarela, S.M. Paulin

Research output: Contribution to journalArticleScientificpeer-review

6 Citations (Scopus)

Abstract

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.
Original languageEnglish
Pages (from-to)49-62
Number of pages14
JournalQuality Assurance and Safety of Crops and Foods
Volume5
DOIs
Publication statusPublished - 2013
MoE publication typeA1 Journal article-refereed

Fingerprint

Campylobacter
microorganisms
Polymerase Chain Reaction
assays
Sensitivity and Specificity
Campylobacter coli
Larus
Campylobacter jejuni
taxonomy

Keywords

  • campylobacter coli
  • campylobacter jejuni
  • identification
  • PCR
  • revalidation

Cite this

On, S.L.W. ; Brandt, S.M. ; Cornelius, A.J. ; Fusco, V. ; Quero, G.M. ; Mackiw, E. ; Houf, K. ; Bilbao, A. ; Diaz, A.I. ; Benejat, L. ; Megraud, F. ; Collins-Emerson, J. ; French, N.P. ; Gotcheva, V. ; Angelov, A. ; Alakomi, Hanna-Leena ; Saarela, M. ; Paulin, S.M. / PCR revisited : A case for revalidation of PCR assays for microorganisms using identification of Campylobacter species as an exemplar. In: Quality Assurance and Safety of Crops and Foods. 2013 ; Vol. 5. pp. 49-62.
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On, SLW, Brandt, SM, Cornelius, AJ, Fusco, V, Quero, GM, Mackiw, E, Houf, K, Bilbao, A, Diaz, AI, Benejat, L, Megraud, F, Collins-Emerson, J, French, NP, Gotcheva, V, Angelov, A, Alakomi, H-L, Saarela, M & Paulin, SM 2013, 'PCR revisited: A case for revalidation of PCR assays for microorganisms using identification of Campylobacter species as an exemplar', Quality Assurance and Safety of Crops and Foods, vol. 5, pp. 49-62. https://doi.org/10.3920/QAS2012.0158

PCR revisited : A case for revalidation of PCR assays for microorganisms using identification of Campylobacter species as an exemplar. / On, S.L.W. (Corresponding Author); Brandt, S.M.; Cornelius, A.J.; Fusco, V.; Quero, G.M.; Mackiw, E.; Houf, K.; Bilbao, A.; Diaz, A.I.; Benejat, L.; Megraud, F.; Collins-Emerson, J.; French, N.P.; Gotcheva, V.; Angelov, A.; Alakomi, Hanna-Leena; Saarela, M.; Paulin, S.M.

In: Quality Assurance and Safety of Crops and Foods, Vol. 5, 2013, p. 49-62.

Research output: Contribution to journalArticleScientificpeer-review

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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

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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

VL - 5

SP - 49

EP - 62

JO - Quality Assurance and Safety of Crops and Foods

JF - Quality Assurance and Safety of Crops and Foods

SN - 1757-8361

ER -