title: Rapid HLA-B27 screening with real-time TaqMan PCR: a clinical validation in the Dutch population.
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.
authors: Department of Clinical Chemistry, Diakonessenhuis, Utrecht, The Netherlands.
authors: Elianne A
authors: EA
authors: Roelandse-Koop
authors: Bert
authors: B
authors: Buisman
authors: Erik J
authors: EJ
authors: van Hannen
authors: Anneke
authors: A
authors: van der Zee
authors: Wouter
authors: W
authors: Kortlandt
authors: Mirjam H A
authors: MH
authors: Hermans
authors: Arend-Jan
authors: AJ
authors: van Houte
authors: Rianne
authors: R
authors: van Rhee-Luderer
mesh_terms: D000483:Alleles; D005091:Exons; D020022:Genetic Predisposition to Disease; D015796:HLA-B27 Antigen / Q000235:genetics*; D006801:Humans; D009426:Netherlands; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013167:Spondylitis, Ankylosing / Q000235:genetics; D044465:White People / Q000235:genetics*
chemical_list: C513088:HLA-B*27:07 antigen; D015796:HLA-B27 Antigen