PMID- 19082079
OWN - NLM
STAT- MEDLINE
DCOM- 20090611
LR  - 20131121
IS  - 1364-5528 (Electronic)
IS  - 0003-2654 (Linking)
VI  - 133
IP  - 12
DP  - 2008 Dec
TI  - A versatile method for quantification of DNA and PCR products based on 
      time-resolved EuIII luminescence.
PG  - 1749-56
LID - 10.1039/b807959e [doi]
AB  - A versatile and robust method for the determination of DNA and PCR products (<500 
      bp) is presented, based on a mix of an Eu(III) chelate and acridine orange (AO). 
      The nucleic acid selective stains acridine orange (AO) and ethidium bromide (EB) 
      quench the luminescence of the bimetallic [Eu(2)(L(C2))(3)] and of other 
      monometallic chelates such as the macrocyclic complex [Eu(L(kel))], even at very 
      low molar ratios. Stern-Volmer plots of the metal-centered emission intensities 
      (F(0)/F) and Eu((5)D(0)) lifetimes (tau(0)/tau) show the AO quenching being 
      purely dynamic with K(D) = 6.7 x 10(5) M(-1) for [Eu(2)(L(C2))(3)] and 1.6 x 
      10(6) M(-1) for [Eu(L(kel))], and bimolecular rate constants k(q) = 2.7 x 10(8) 
      M(-1) s(-1) and 3.4 x 10(9) M(-1) s(-1), respectively. On the other hand, EB 
      quenching is due to both dynamic and static mechanisms. In the presence of 
      various types of DNA > 0.1 ng microL(-1) (dsDNA, ssDNA or circular DNA), the 
      quenched luminescence is reinstated, AO and EB intercalating into DNA, which 
      removes the interaction with the Eu(III) complexes. The best results are obtained 
      with [Eu(2)(L(C2))(3)]/AO with detections limits in the range 0.18-0.66 ng 
      microL(-1); detection limits for the [Eu(L(kel))]/AO system are slightly larger; 
      simpler monometallic Eu(III) complexes with dipicolinate derivatives do not 
      follow suit in that they decompose in the presence of DNA. The Eu(III)/AO method 
      is shown to be pH insensitive in the range 3-10; furthermore it is essentially 
      insensitive to 1000-fold excesses of potential interfering substances, e.g. BSA, 
      glucose, chelating agents and anions, alkaline earth and transition metal 
      cations, variations in luminescence intensity being < 5%, (10 analytes) or 5-10% 
      (4 analytes); only Co(II) and Cu(II) interfere substantially.
FAU - Song, Bo
AU  - Song B
AD  - Laboratory of Lanthanide Supramolecular Chemistry, Ecole Polytechnique Federale 
      de Lausanne, Lausanne, Switzerland.
FAU - Vandevyver, Caroline D B
AU  - Vandevyver CD
FAU - Deiters, Emmanuel
AU  - Deiters E
FAU - Chauvin, Anne-Sophie
AU  - Chauvin AS
FAU - Hemmila, Ilkka
AU  - Hemmila I
FAU - Bunzli, Jean-Claude G
AU  - Bunzli JC
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20080905
PL  - England
TA  - Analyst
JT  - The Analyst
JID - 0372652
RN  - 0 (Chelating Agents)
RN  - 444W947O8O (Europium)
RN  - 9007-49-2 (DNA)
RN  - EN464416SI (Ethidium)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange
MH  - Chelating Agents
MH  - DNA/*analysis
MH  - Ethidium
MH  - Europium/*chemistry
MH  - Luminescence
MH  - Photochemical Processes
MH  - Polymerase Chain Reaction/methods
EDAT- 2008/12/17 09:00
MHDA- 2009/06/12 09:00
CRDT- 2008/12/17 09:00
PHST- 2008/12/17 09:00 [entrez]
PHST- 2008/12/17 09:00 [pubmed]
PHST- 2009/06/12 09:00 [medline]
AID - 10.1039/b807959e [doi]
PST - ppublish
SO  - Analyst. 2008 Dec;133(12):1749-56. doi: 10.1039/b807959e. Epub 2008 Sep 5.