Clinical Chemistry AACC Online Job Center
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Clinical Chemistry 49: 782-791, 2003; 10.1373/49.5.782
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Submit an electronic Letter to
the Editor about this paper
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (13)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Raggi, C. C.
Right arrow Articles by Orlando, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Raggi, C. C.
Right arrow Articles by Orlando, C.
Related Collections
Right arrow Molecular Diagnostics and Genetics
Right arrow Laboratory Management
(Clinical Chemistry. 2003;49:782-791.)
© 2003 American Association for Clinical Chemistry, Inc.

External Quality Assurance Program for PCR Amplification of Genomic DNA: An Italian Experience

Claudia Casini Raggi1, Pamela Pinzani1, Angelo Paradiso2, Mario Pazzagli1 and Claudio Orlando1,a

1 Clinical Biochemistry Unit, Department of Clinical Physiopathology, University of Florence, viale Pieraccini 6, 50139 Florence, Italy.

2 Clinical Experimental Oncology Laboratory, National Cancer Institute, 70126 Bari, Italy.

aAuthor for correspondence. Fax 39-055-4271-413; e-mail c.orlando{at}dfc.unifi.it.

Background: External quality assurance (EQA) programs for diagnostic tests based on nucleic acid amplification have not been widely implemented in clinical laboratories and remain limited to few tests. Development of specific EQA programs based on application-based proficiency testing for any diagnostic molecular target is challenging. Development of EQA trials based on methodologic proficiency testing and directed to the evaluation of analytical aspects common to the majority of PCR-based tests may be valuable.

Methods: We developed an EQA program for evaluation of DNA extraction and amplification and analysis of products after PCR. Participants received a package containing primers and reference materials to evaluate three specific controls for, respectively, DNA extraction (quality and quantity), PCR performance (specificity and efficiency), and interpretation of results after electrophoresis. Each participant was asked to return to the organizers a form with their numerical results and an aliquot of all amplified samples for joint evaluation.

Results: Results varied in all phases of the experimental procedure: preamplification, amplification, and post-PCR interpretation. To give a general estimation on the quality of performances for each laboratory, we designed a score scheme in which the results of any specific action were evaluated on the basis of the distribution around the median consensus values. The maximum possible score was 84. On the basis of total score obtained by each laboratory, we created a qualitative ranking list that provided the final interpretation of results as excellent (>63 points; n = 4 laboratories), good (53–63 points; n = 13), sufficient (42–52 points; n = 15), poor (31–41 points; n = 3), and not acceptable (<31 points; n = 4).

Conclusions: This survey demonstrates the importance of EQA trials based on methodologic proficiency testing directed to evaluation of analytical aspects common to the majority of PCR-based tests.




The following articles in journals at HighWire Press have cited this article:


Home page
 Clin. Chem.Home page
C. Orlando, P. Verderio, R. Maatman, J. Danneberg, S. Ramsden, M. Neumaier, D. Taruscio, V. Falbo, R. Jansen, C. Casini-Raggi, et al.
EQUAL-qual: A European Program for External Quality Assessment of Genomic DNA Extraction and PCR Amplification
Clin. Chem., July 1, 2007; 53(7): 1349 - 1357.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
S. J. Patton, A. J. Wallace, and R. Elles
Benchmark for Evaluating the Quality of DNA Sequencing: Proposal from an International External Quality Assessment Scheme
Clin. Chem., April 1, 2006; 52(4): 728 - 736.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
S. Raja, J. Ching, L. Xi, S. J. Hughes, R. Chang, W. Wong, W. McMillan, W. E. Gooding, K. S. McCarty Jr, M. Chestney, et al.
Technology for Automated, Rapid, and Quantitative PCR or Reverse Transcription-PCR Clinical Testing
Clin. Chem., May 1, 2005; 51(5): 882 - 890.
[Abstract] [Full Text] [PDF]

Home page
 Endocr Relat CancerHome page
V. Cappelletti, L. Celio, E. Bajetta, A. Allevi, R. Longarini, P. Miodini, R. Villa, A. Fabbri, L. Mariani, R. Giovanazzi, et al.
Prospective evaluation of estrogen receptor-{beta} in predicting response to neoadjuvant antiestrogen therapy in elderly breast cancer patients
Endocr. Relat. Cancer, December 1, 2004; 11(4): 761 - 770.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
L. Birch, C. A. English, M. Burns, and J. T. Keer
Generic Scheme for Independent Performance Assessment in the Molecular Biology Laboratory
Clin. Chem., September 1, 2004; 50(9): 1553 - 1559.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
C. S. Richards and W. W. Grody
Alternative Approaches to Proficiency Testing in Molecular Genetics
Clin. Chem., May 1, 2003; 49(5): 717 - 718.
[Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2003 by the American Association for Clinical Chemistry.