PMID- 15336784
OWN - NLM
STAT- MEDLINE
DCOM- 20041019
LR  - 20061115
IS  - 0198-8859 (Print)
IS  - 0198-8859 (Linking)
VI  - 65
IP  - 8
DP  - 2004 Aug
TI  - HLA-B27 genotyping by fluorescent resonance emission transfer (FRET) probes in 
      real-time PCR.
PG  - 826-38
AB  - Several polymerase chain reaction (PCR)-based human leukocyte antigen (HLA) 
      genotyping methods are in use, but none is fully satisfactory. The introduction 
      of real-time PCR (rt-PCR) with fluorescence resonance energy transfer (FRET) 
      probes provides a powerful tool to overcome the drawbacks of current methods such 
      as the long processing time and the requirement for post-PCR manual procedures. 
      Here we present evidence that the FRET-fluorotyping principle may resolve HLA-B27 
      variants, providing a higher resolution in less time than the techniques 
      currently in use. The protocol uses between one and three consecutive 
      amplification reactions depending on the resolution required. The first reaction, 
      aimed at detecting HLA-B27-positive samples, uses beta-globin coamplification as 
      control. The second reaction, aimed at resolving most frequent B27 alleles, uses 
      two hybridization probes whose melting temperatures curves allow the 
      classification of HLA-B27 alleles into eight groups. By adding a third reaction, 
      even the rarest alleles associated and not associated to ankylosing spondylitis 
      (AS) may be discriminated. The technique was blindly tested on 60 samples from 
      individuals previously typed and confirmed by standard PCR sequence-specific 
      oligoprobes-PCR sequence and PCR-based typing PCR-SBT (30 B27+, 30 non-B27). 
      There was a complete concordance rate, thus confirming the potential of this new 
      technique for clinical HLA-B27 typing and for HLA typing in general.
FAU - Faner, Rosa
AU  - Faner R
AD  - Laboratory of Immunobiology Research and Applications to Diagnosis (LIRAD), 
      Serveis Sanitaris de Referencia-Centre de Transfusio i Banc de Teixits 
      (SSR-CTBT), Badalona, Barcelona, Spain.
FAU - Casamitjana, Natalia
AU  - Casamitjana N
FAU - Colobran, Roger
AU  - Colobran R
FAU - Ribera, Anna
AU  - Ribera A
FAU - Pujol-Borrell, Ricardo
AU  - Pujol-Borrell R
FAU - Palou, Eduard
AU  - Palou E
FAU - Juan, Manel
AU  - Juan M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Hum Immunol
JT  - Human immunology
JID - 8010936
RN  - 0 (HLA-B27 Antigen)
SB  - IM
MH  - Cell Line, Transformed
MH  - Fluorescence Resonance Energy Transfer/*methods
MH  - Gene Frequency
MH  - Genotype
MH  - HLA-B27 Antigen/classification/*genetics
MH  - Histocompatibility Testing/*methods
MH  - Humans
MH  - Polymerase Chain Reaction/*methods
EDAT- 2004/09/01 05:00
MHDA- 2004/10/20 09:00
CRDT- 2004/09/01 05:00
PHST- 2004/03/01 00:00 [received]
PHST- 2004/05/10 00:00 [revised]
PHST- 2004/05/19 00:00 [accepted]
PHST- 2004/09/01 05:00 [pubmed]
PHST- 2004/10/20 09:00 [medline]
PHST- 2004/09/01 05:00 [entrez]
AID - S0198-8859(04)00125-9 [pii]
AID - 10.1016/j.humimm.2004.05.009 [doi]
PST - ppublish
SO  - Hum Immunol. 2004 Aug;65(8):826-38. doi: 10.1016/j.humimm.2004.05.009.