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The Italian external quality assessment for RAS testing in colorectal carcinoma identifies methods-related inter-laboratory differences.

Normanno N, Pinto C, Castiglione F, Fenizia F, Barberis M, Marchetti A, Fontanini G, De Rosa G, Taddei GL - J Transl Med (2015)

Bottom Line: In the first round 36% of the 88 participants failed, with 23 centers having at least one false positive or false negative while 9 centers did not meet the deadline.The genotyping error rate was higher when Sanger sequencing was employed for testing as compared with pyrosequencing (3 vs 1.3%; p = 0.01; Pearson Chi Square test).The results of the Italian RAS EQA scheme indicate that the mutational analyses are performed with good quality in many Italian centers, although significant differences in the methods used were highlighted.

View Article: PubMed Central - PubMed

Affiliation: Cell Biology and Biotherapy Unit, Istituto Nazionale per lo Studio e la Cura dei Tumori "Fondazione Giovanni Pascale"-IRCCS, Via M. Semmola, 80131, Naples, Italy. nicnorm@yahoo.com.

ABSTRACT

Background: In 2014 the European Medicines Agency included exon 2, 3 and 4 KRAS and NRAS testing for the selection of metastatic colorectal cancer (mCRC) patients eligible for the therapy with anti-EGFR monoclonal antibodies. The Italian Association of Medical Oncology (AIOM) and the Italian Society of Pathology and Cytology (SIAPEC) organized an external quality assessment (EQA) scheme for CRC to evaluate inter-laboratory consistency and to ensure standardization of the results in the transition from KRAS to all-RAS testing.

Methods: Ten formalin fixed paraffin embedded specimens including KRAS/NRAS (exons 2, 3, 4) and BRAF (codon 600) mutations were validated by three referral laboratories and sent to 88 participant centers. Molecular pathology sample reports were also requested to each laboratory. A board of assessors from AIOM and SIAPEC evaluated the results according to a predefined scoring system. The scheme was composed of two rounds.

Results: In the first round 36% of the 88 participants failed, with 23 centers having at least one false positive or false negative while 9 centers did not meet the deadline. The genotyping error rate was higher when Sanger sequencing was employed for testing as compared with pyrosequencing (3 vs 1.3%; p = 0.01; Pearson Chi Square test). In the second round, the laboratories improved their performance, with 23/32 laboratories passing the round. Overall, 79/88 participants passed the RAS EQA scheme. Standardized Human Genome Variation Society nomenclature was incorrectly used to describe the mutations identified and relevant variations were noticed in the genotype specification.

Conclusion: The results of the Italian RAS EQA scheme indicate that the mutational analyses are performed with good quality in many Italian centers, although significant differences in the methods used were highlighted. The relatively high number of centers failing the first round underlines the fundamental role in continued education covered by EQA schemes.

No MeSH data available.


Related in: MedlinePlus

Results of the Italian external quality assessment for all-RAS and BRAF testing in colorectal carcinoma
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Fig2: Results of the Italian external quality assessment for all-RAS and BRAF testing in colorectal carcinoma

Mentions: Eventually, 79 of 88 participants obtained a score ≥18 in the first or in the second round and passed the RAS EQA scheme, including 2 foreign centers (Fig. 2). The list of the laboratories that passed the scheme is published on the web site of AIOM and SIAPEC (http://www.aiom.it; http://www.siapec.it).Fig. 2


The Italian external quality assessment for RAS testing in colorectal carcinoma identifies methods-related inter-laboratory differences.

Normanno N, Pinto C, Castiglione F, Fenizia F, Barberis M, Marchetti A, Fontanini G, De Rosa G, Taddei GL - J Transl Med (2015)

Results of the Italian external quality assessment for all-RAS and BRAF testing in colorectal carcinoma
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4557483&req=5

Fig2: Results of the Italian external quality assessment for all-RAS and BRAF testing in colorectal carcinoma
Mentions: Eventually, 79 of 88 participants obtained a score ≥18 in the first or in the second round and passed the RAS EQA scheme, including 2 foreign centers (Fig. 2). The list of the laboratories that passed the scheme is published on the web site of AIOM and SIAPEC (http://www.aiom.it; http://www.siapec.it).Fig. 2

Bottom Line: In the first round 36% of the 88 participants failed, with 23 centers having at least one false positive or false negative while 9 centers did not meet the deadline.The genotyping error rate was higher when Sanger sequencing was employed for testing as compared with pyrosequencing (3 vs 1.3%; p = 0.01; Pearson Chi Square test).The results of the Italian RAS EQA scheme indicate that the mutational analyses are performed with good quality in many Italian centers, although significant differences in the methods used were highlighted.

View Article: PubMed Central - PubMed

Affiliation: Cell Biology and Biotherapy Unit, Istituto Nazionale per lo Studio e la Cura dei Tumori "Fondazione Giovanni Pascale"-IRCCS, Via M. Semmola, 80131, Naples, Italy. nicnorm@yahoo.com.

ABSTRACT

Background: In 2014 the European Medicines Agency included exon 2, 3 and 4 KRAS and NRAS testing for the selection of metastatic colorectal cancer (mCRC) patients eligible for the therapy with anti-EGFR monoclonal antibodies. The Italian Association of Medical Oncology (AIOM) and the Italian Society of Pathology and Cytology (SIAPEC) organized an external quality assessment (EQA) scheme for CRC to evaluate inter-laboratory consistency and to ensure standardization of the results in the transition from KRAS to all-RAS testing.

Methods: Ten formalin fixed paraffin embedded specimens including KRAS/NRAS (exons 2, 3, 4) and BRAF (codon 600) mutations were validated by three referral laboratories and sent to 88 participant centers. Molecular pathology sample reports were also requested to each laboratory. A board of assessors from AIOM and SIAPEC evaluated the results according to a predefined scoring system. The scheme was composed of two rounds.

Results: In the first round 36% of the 88 participants failed, with 23 centers having at least one false positive or false negative while 9 centers did not meet the deadline. The genotyping error rate was higher when Sanger sequencing was employed for testing as compared with pyrosequencing (3 vs 1.3%; p = 0.01; Pearson Chi Square test). In the second round, the laboratories improved their performance, with 23/32 laboratories passing the round. Overall, 79/88 participants passed the RAS EQA scheme. Standardized Human Genome Variation Society nomenclature was incorrectly used to describe the mutations identified and relevant variations were noticed in the genotype specification.

Conclusion: The results of the Italian RAS EQA scheme indicate that the mutational analyses are performed with good quality in many Italian centers, although significant differences in the methods used were highlighted. The relatively high number of centers failing the first round underlines the fundamental role in continued education covered by EQA schemes.

No MeSH data available.


Related in: MedlinePlus