3rd meeting of young researchers at UP 1 - IJUP - Universidade do ...

Crossing between PCR and HACCP: a promising new approach?
A. Oliveira 1,2 , M.S.J. Nascimento 2,3 , R. Medeiros 1
1 Molecular Oncology Unit, Portuguese Institute of Oncology Francisco Gentil, Laboratórios
Piso 4, Rua Dr. António Bernardino Almeida, Porto, 4200-072, Portugal
2 Department of Microbiology, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha,
164, 4050-047 Porto, Portugal
3 CEQUIMED-UP, Faculty of Pharmacy, University of Porto, Portugal
In vitro amplification of nucleic acids using PCR has become a powerful diagnostic tool.
However, the lack of officially approved standardized regulations and instructions has been
pointed as one of the disadvantages, as well as the difficulty to control all the variables, which
can compromise the quality of the results. So, it is necessary to introduce a quality control plan
and management system, in order to achieve the new universal trend of quality assurance [1,2].
HACCP is a preventive approach based on a scientific and logical system designed to identify
and evaluate specific hazards at all stages of the process. Despite the use of HACCP has not
yet expanded to the genetic laboratory, its foundation can be adapted and implemented in
different sectors, with high probability of success. [3,4]
The aim of this work is to present a proposal for assessing internal quality of the technique of
PCR-RFLP and Real-Time PCR, used daily in the laboratory of Molecular Oncology of IPO-
Porto. Through a careful observation of the laboratory and its collaborators, and using the
fundamentals of quality management system HACCP, the flowcharts were developed for the
techniques, as well as tables for the identification of hazards, critical control points, control and
corrective measures, bringing together all the actions to be undertaken during the technique
and problem-solving. Some criteria were also adjusted for evaluating the various sectors of the
laboratory, and used for daily observation. These may be implemented as future measures of
internal evaluation of the site, its organization and collaborators.
Using the fundamentals of HACCP, the criteria and observation, it was concluded that the
main requirements of work are generally satisfied, but needed a protocol to evaluate the
internal quality of the various steps, in addition to the results. A plan of internal quality control
was prepared, to experiment, improve and implement in the near future making possible the
crossing between PCR and HACCP.
References:
[1] Williams L.O., Cole E.C., Lubin I.M., Iglesias N.I., Jordan R.L. and Elliott L.E. (2003),
Quality assurance in human molecular genetics testing, Arch Pathol Lab Med, 127 (10), 1353-
1358.
[2] Bellissimo D.B. (2007), Practice guidelines and proficiency testing for molecular assays,
Transfusion, 47 (Suppl 1), 79S-84S.
[3] Stroobants A.K., Goldschmidt H.M.J. and Plebani M. (2003), Errorbudget calculations in
laboratory medicine: linking the concepts of biological variation and allowable medical
errors, Clin Chim Acta, 333 (2), 169-176.
[4] Lippi G., Guidi G.C., Mattiuzzi C. and Plebani M. (2006), Preanalytical variability: the
dark side of the moon in laboratory testing, Clin Chem Lab Med, 44 (4), 358-365.
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