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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

12 Issues per year


IMPACT FACTOR increased in 2015: 3.017
Rank 5 out of 30 in category Medical Laboratory Technology in the 2014 Thomson Reuters Journal Citation Report/Science Edition

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Online
ISSN
1437-4331
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Volume 49, Issue 12 (Dec 2011)

Issues

Rapid HLA-B27 screening with real-time TaqMan PCR: a clinical validation in the Dutch population

Elianne A. Roelandse-Koop
  • Department of Clinical Chemistry, Diakonessenhuis, Utrecht, The Netherlands
  • Current address: Free University Medical Centre, Amsterdam, The Netherlands.
/ Bert Buisman
  • Department of Clinical Chemistry, Diakonessenhuis, Utrecht, The Netherlands
/ Erik J. van Hannen
  • Department of Medical Microbiology and Immunology, St. Antonius Hospital, Nieuwegein, The Netherlands
/ Anneke van der Zee
  • Laboratory of Medical Microbiology, St. Elisabeth Hospital, Tilburg, The Netherlands
  • Current address: Maasstad Hospital, Rotterdam, The Netherlands.
/ Wouter Kortlandt
  • Department of Clinical Chemistry, Diakonessenhuis, Utrecht, The Netherlands
/ Mirjam H.A. Hermans
  • Molecular Diagnostics, Jeroen Bosch Hospital, ‘s Hertogenbosch, The Netherlands
/ Arend-Jan van Houte
  • Department of Clinical Chemistry, Diakonessenhuis, Utrecht, The Netherlands
  • Department of Medical Microbiology and Immunology, St. Antonius Hospital, Nieuwegein, The Netherlands
  • Department of Medical Microbiology and Immunology, Diakonessenhuis, Utrecht, The Netherlands
  • Email:
/ Rianne van Rhee-Luderer
  • Unit Molecular Diagnostics, Diakonessenhuis, Utrecht, The Netherlands
  • Current address: Centre for Infectious Diseases Friesland, Leeuwarden, The Netherlands and Molecular Diagnostic Centre Friesland, Leeuwarden, The Netherlands.
Published Online: 2011-09-06 | DOI: https://doi.org/10.1515/CCLM.2011.252

Abstract

Background: Human leukocyte antigen B27 (HLA-B27) is strongly associated with ankylosing spondylitis. The B27 allele is present in 90% of patients with this disease, whereas it is present in only 9% of Caucasians. Molecular detection of HLA-B27 is traditionally based on allele specific amplification of exon 2 (Olerup method) or exon 3 (Dominguez method) by PCR, followed by gel analysis.

Methods: We developed a real-time TaqMan PCR based on the Dominguez method with a β-Globin PCR as internal control.

Results: A total of 544 clinical samples were used to compare the real-time TaqMan PCR with the traditional Dominguez PCR, the traditional Olerup PCR and a commercial Olerup based HLA-B27 detection kit (Olerup SSPTM HLA-B27, GenoVision). While 542 samples gave concordant results, two samples showed discrepancies and were further analyzed. One sample that showed a discrepancy was negative with the traditional Olerup method and positive with the three other procedures. Sequencing analysis showed the presence of HLA-B*2712 in this sample. The other sample, positive with both Olerup based PCRs and negative with both Dominguez based methods, turned out to be positive for HLA-B*2707 by sequence analysis.

Conclusions: With a correct result for 543 out of 544 samples (99.8%), we consider our real-time HLA-B27 PCR is a reliable method to detect HLA-B27 in the Dutch population, with reduced hands-on time and contamination risk compared to traditional PCR methods.

Keywords: ankylosing spondylitis; Dutch population; HLA-B27; real-time PCR; TaqMan

About the article

Corresponding author: Dr. Arend-Jan van Houte, Bosboomstraat 1, 3582 KE Utrecht, The Netherlands Phone: +31-882506697, Fax: +31-882506695


Received: 2010-11-29

Accepted: 2011-08-01

Published Online: 2011-09-06

Published in Print: 2011-12-01


Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2011.252. Export Citation

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