PMID- 25120130
OWN - NLM
STAT- MEDLINE
DCOM- 20151001
LR  - 20191210
IS  - 1096-0309 (Electronic)
IS  - 0003-2697 (Linking)
VI  - 465
DP  - 2014 Nov 15
TI  - Closed-tube human leukocyte antigen DQA1 *05 genotyping assay based on switchable 
      lanthanide luminescence probes.
PG  - 6-11
LID - S0003-2697(14)00331-5 [pii]
LID - 10.1016/j.ab.2014.07.029 [doi]
AB  - Genotyping in closed tube is commonly performed using polymerase chain reaction 
      (PCR) amplification and allele-specific oligonucleotide probes using fluorescence 
      resonance energy transfer (FRET). Here we introduce a homogeneous human leukocyte 
      antigen (HLA)-DQA1 *05 end-point PCR assay based on switchable lanthanide 
      luminescence probe technology and a simple dried blood sample preparation. The 
      switchable probe technology is based on two non-luminescent oligonucleotide 
      probes: one carrying a non-luminescent lanthanide chelate and the other carrying 
      a light-absorbing antenna ligand. Hybridization of the probes in adjacent 
      positions to the target DNA leads to the formation of a highly luminescent 
      lanthanide chelate complex by self-assembly of the reporter molecules. 
      Performance of the HLA-DQA1 *05 assay was evaluated by testing blood samples 
      collected on sample collection cards and was prepared by lysing the punched 
      samples (3-mm discs) using alkaline reaction conditions and high temperature. 
      Testing of 147 blood samples yielded 100% correlation to the heterogeneous DELFIA 
      technology-based reference assay. Genotyping requires carefully designed probe 
      sequences able to discriminate matched and mismatched target sequences by 
      hybridization. Furthermore, definite genotype discrimination was achieved because 
      inherently non-luminescent switchable probes together with time-resolved 
      measurement mode led to very low background signal level and, therefore, very 
      high signal differences averaging 54-fold between DQA1 *05 and other alleles.
CI  - Copyright (c) 2014 Elsevier Inc. All rights reserved.
FAU - Lehmusvuori, Ari
AU  - Lehmusvuori A
AD  - Department of Biochemistry/Biotechnology, University of Turku, 20520 Turku, 
      Finland. Electronic address: ari.lehmusvuori@utu.fi.
FAU - Kiviniemi, Minna
AU  - Kiviniemi M
AD  - Immunogenetics Laboratory and MediCity Research Laboratory, University of Turku, 
      20520 Turku, Finland.
FAU - Ilonen, Jorma
AU  - Ilonen J
AD  - Immunogenetics Laboratory and MediCity Research Laboratory, University of Turku, 
      20520 Turku, Finland; Department of Clinical Microbiology, University of Eastern 
      Finland, FI-70211 Kuopio, Finland.
FAU - Soukka, Tero
AU  - Soukka T
AD  - Department of Biochemistry/Biotechnology, University of Turku, 20520 Turku, 
      Finland.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20140810
PL  - United States
TA  - Anal Biochem
JT  - Analytical biochemistry
JID - 0370535
RN  - 0 (DNA Probes)
RN  - 0 (HLA-DQ alpha-Chains)
RN  - 0 (HLA-DQA1 antigen)
RN  - 444W947O8O (Europium)
SB  - IM
MH  - DNA Probes/*chemistry/genetics
MH  - Europium/*chemistry
MH  - Fluorescence Resonance Energy Transfer/*methods
MH  - Genotyping Techniques/*methods
MH  - HLA-DQ alpha-Chains/*genetics
MH  - Humans
MH  - Luminescent Measurements/*methods
OTO - NOTNLM
OT  - Genotyping
OT  - Lanthanide chelate
OT  - PCR
OT  - Switchable sensor
EDAT- 2014/08/15 06:00
MHDA- 2015/10/02 06:00
CRDT- 2014/08/15 06:00
PHST- 2014/05/09 00:00 [received]
PHST- 2014/07/30 00:00 [revised]
PHST- 2014/07/31 00:00 [accepted]
PHST- 2014/08/15 06:00 [entrez]
PHST- 2014/08/15 06:00 [pubmed]
PHST- 2015/10/02 06:00 [medline]
AID - S0003-2697(14)00331-5 [pii]
AID - 10.1016/j.ab.2014.07.029 [doi]
PST - ppublish
SO  - Anal Biochem. 2014 Nov 15;465:6-11. doi: 10.1016/j.ab.2014.07.029. Epub 2014 Aug 
      10.