PMID- 15620466
OWN - NLM
STAT- MEDLINE
DCOM- 20051011
LR  - 20061115
IS  - 0198-8859 (Print)
IS  - 0198-8859 (Linking)
VI  - 66
IP  - 1
DP  - 2005 Jan
TI  - Development of a new HLA-DRB real-time PCR typing method.
PG  - 85-91
AB  - Polymerase chain reaction (PCR)-based human leukocyte antigen (HLA) typing 
      methods currently used in most histocompatibility laboratories, such as 
      PCR-sequence-specific primers (PCR-SSP) and PCR-sequence-specific oligonucleotide 
      probes (PCR-SSO), are time-consuming and are at risk of contamination during the 
      post-PCR process. The aim of this study was to develop a real-time PCR 
      (rtPCR)-based HLA-DRB1 and -DRB3/4/5 low-medium resolution typing method to avoid 
      these problems. This new method combined the use of specific primers and probes 
      for HLA-DRB alleles. One pair of DRB gene primers and two DRB-specific probes 
      (FAM and VIC) were used per reaction in each of a set of 16 PCR reaction tubes. 
      To provide an internal positive control, each tube also contained a pair of 
      primers and a TET probe for glyceraldehyde phosphate dehydrogenase. This allowed 
      a very significant reduction in the number of reactions and the processing time, 
      whereas typing resolution increased. After successful testing on 100 samples, the 
      technique was validated in 200 clinical samples that had previously been typed 
      for HLA-DRB using a standard PCR-based method. Identical results were obtained 
      with all samples. This new method also reduced ambiguous results and was faster 
      and less cumbersome than currently used PCR-SSP or PCR-SSO techniques.
FAU - Casamitjana, Natalia
AU  - Casamitjana N
AD  - Laboratori d'Immunobiologia per a la Recerca i Aplicacions Diagnostiques, Serveis 
      Sanitaris de Referencia--Centre de Transfusio i Banc de Teixits, Barcelona, 
      Spain.
FAU - Faner, Rosa
AU  - Faner R
FAU - Santamaria, Albert
AU  - Santamaria A
FAU - Colobran, Roger
AU  - Colobran R
FAU - Ribera, Anna
AU  - Ribera A
FAU - Pujol-Borrell, Ricardo
AU  - Pujol-Borrell R
FAU - Juan, Manel
AU  - Juan M
FAU - Palou, Eduard
AU  - Palou E
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 (DNA Primers)
RN  - 0 (HLA-DR Antigens)
RN  - EC 2.7.7.- (Taq Polymerase)
SB  - IM
MH  - *Alleles
MH  - DNA Primers
MH  - HLA-DR Antigens/*analysis/genetics/immunology
MH  - Histocompatibility Testing/methods
MH  - Humans
MH  - Organ Transplantation
MH  - Polymerase Chain Reaction/*methods
MH  - Taq Polymerase
EDAT- 2004/12/29 09:00
MHDA- 2005/10/12 09:00
CRDT- 2004/12/29 09:00
PHST- 2004/04/26 00:00 [received]
PHST- 2004/07/29 00:00 [revised]
PHST- 2004/08/06 00:00 [accepted]
PHST- 2004/12/29 09:00 [pubmed]
PHST- 2005/10/12 09:00 [medline]
PHST- 2004/12/29 09:00 [entrez]
AID - S0198-8859(04)00585-3 [pii]
AID - 10.1016/j.humimm.2004.08.178 [doi]
PST - ppublish
SO  - Hum Immunol. 2005 Jan;66(1):85-91. doi: 10.1016/j.humimm.2004.08.178.