THE EQUAL-qual Programme

1. Objectives of the EQUAL-qual programme

Today the polymerase chain reaction (PCR) is still one of the most powerful techniques used in many laboratories for molecular biology. PCR is extensively used in diverse fields of HealthCare as pathology, clinical chemistry, microbiology, and cytogenetics. However, external quality control programmes (EQA) designed to monitor the performance of PCR and PCR-related assays have been very limited in Europe. The EQUAL-qual program permits the evaluation of several analytical aspects common to the majority of the molecular tests based on qualitative PCR.

The program has been designed to evaluate PCR assays at three control levels:

1. DNA extraction (quality and quantity);
2. PCR performances (specificity and efficiency);
3. Interpretation of the results after electrophoresis.

2. Participant selection

Participation is open to all laboratories carrying out PCR based assays within the EU. Clinical laboratories of different medical professions (e.g. Clinical Chemistry, Pathology, Human Genetics, Microbiology, Forensics) are especially invited to join the programme. In the EQUAL-qual program, up to 423 Laboratories can participate (see the geographical distribution of participants at Description of the EQUAL Programmes). Participation is free.

3. What do you need to participate?

Participating laboratories need all the facilities to perform a PCR, like a thermal cycler, a spectrophotometer (UV), and a system for post-PCR analysis (like a submarine gel-electrophoresis system).
You will be requested to use your thermal cycler/s and the reagents commonly in use in your laboratory. Also instruments to record your PCR results (self-developing photos or image analysers) are necessary.

4. What will you find in the provided EQUAL-qual kit? Which actions do you have to do?

This is the list of the material included in the EQUAL-qual kit and of the actions to be performed in your laboratory.

Sample/reagent	Concentration	Actions
		If necessary extract DNA and determine: DNA quantity DNA quality (A260/A280 ratio) Report results	PCR with primer- set A + B. Report results	Send PCR products to project coordinator
Sample 1 1200 µL blood (citrate)	Blood contains 4 – 10 109/L leukocytes	X	X	X
Sample 2 1200 µL blood (citrate)	Blood contains 4 – 10 109/L leukocytes	X	X	X
Sample 3 50 µL DNA sample	[DNA] ca. 30 ng/µL	X	X	X
Sample 4 50 µL DNA sample	[DNA] ca. 30 ng/µL	X	X	X
Reagent A Primerset A 40 µL	20 µmol/L
Reagent B Primerset B 40 µL	20 µmol/L

5. Description of the actions

• DNA extraction: by using your own procedure, DNA should be extracted from 100-500 µl of the two provided blood samples.
• DNA quality and quantity: by using spectrophotometric analysis, you should evaluate DNA quality/quantity in four samples: the two blood-derived extracts (1 & 2) and the two pre-extracted DNA samples (3 & 4).
• PCR: by using the two ready-to-use primer sets (Reagent A and Reagent B), you will run a single PCR experiment on samples 1, 2, 3 and 4, for a total of eight different tubes.
• Check the experiment: by using your gel-electrophoresis apparatus, you will verify the results of your PCR amplifications.
• Submit the data: you will be requested to insert in the web-site (www.equal.unifi.it) the answer for each requested information.
• Send an aliquot: an aliquot of your 8 PCR products as well as aliquots of DNA extracted from sample 1 and 2, must be sent by express mail to the EQUAL-qual project management (shipment box enclosed in the kit) for further analysis (please note that the expenses for this expedition must be covered by your institution).
• Find the reports and your score: your performances and those of all participants will be evaluated anonymously and they will be consultable on the web-site (www.equal.unifi.it).
  Individual performance, with the final score and comments will also be sent to participants by mail.

A detailed description of the material enclosed in the EQUAL-qual Kit and of the actions to be performed will be enclosed in the EQUAL-qual Kit.

6. Time Schedule of the survey

Deadline for application	November, 6, 2004
Shipment of the EQUAL-qual kit to participants	starts October 2004 ends December 2004
Phase of analysis in the labs, uploading of the results and shipment of the samples	October, 2004 - February, 2005
Closing of the survey	February 26, 2005
Re-evaluation of materials at the facilities of the organizers	until end of April 2005
Statistical analysis on the Web site	May 2005
Back-Reporting	May 2005
Training Courses	planned in the period July-October 2005

7. Do you want to know more on it?

Previous experiences have been performed at National levels and some of them have been published (see the Links page). Do you need further information? Please send a message to A. Martens by e-mail (a.martens@twenteborg.nl) or by mail to:

A. Martens, MD
Clinical Laboratory
Ziekenhuisgroep Twente
Postbox 7600
7600 SZ Almelo
The Netherlands

8. Do you want to join the EQUAL-qual program?

Please fill your application form by clicking here.

 

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