PMID- 20209349
OWN - NLM
STAT- MEDLINE
DCOM- 20100510
LR  - 20190907
IS  - 1678-9849 (Electronic)
IS  - 0037-8682 (Linking)
VI  - 42
IP  - 6
DP  - 2009 Nov-Dec
TI  - [DNA extraction from coagulated human blood for application in genotyping 
      techniques for human leukocyte antigen and immunoglobulin-like receptors].
PG  - 651-6
LID - S0037-86822009000600008 [pii]
AB  - The objective of this study was to standardize a method for extracting 
      high-quality DNA from samples of coagulated blood. Forty-eight samples of human 
      coagulated blood were used for DNA extraction by means of the EZ-DNA commercial 
      kit (Biological Industries, Beit Haemek, Israel), the Neoscience column kit (One 
      Lambda Inc., San Diego, CA, USA) and a modified salting-out method. Only the 
      salting-out method was able to extract high concentrations of DNA (mean, 180 
      ng/(1/4)microl), which were measured using the Qubit fluorescence detector 
      (Invitrogen, USA). This method enabled amplification of HLA (human leukocyte 
      antigen) genes using the Luminex PCR-SSO (polymerase chain reaction - 
      sequence-specific oligonucleotide) technology, which demands good quality DNA, 
      and amplification of KIR (killer-cell immunoglobulin-like receptor) genes using 
      an in-house PCR-SSP (polymerase chain reaction - sequence-specific primer) 
      technique, which demands a specific concentration of DNA (10 ng/(1/4)microl). We 
      concluded that the modified salting-out technique was very efficient, simple and 
      fast for DNA extraction from human coagulated blood samples, with the aim of 
      genotyping the HLA and KIR genes.
FAU - Cardozo, Daniela Maira
AU  - Cardozo DM
AD  - Laboratorio de Imunogenetica, Departamento de Analises Clinicas, Universidade 
      Estadual de Maringa, Maringa, PR. jelvisentainer@uem.br
FAU - Guelsin, Glaucia Andreia
AU  - Guelsin GA
FAU - Clementino, Samaia Laface
AU  - Clementino SL
FAU - Melo, Fabiano Cavalcante de
AU  - Melo FC
FAU - Braga, Marco Antonio
AU  - Braga MA
FAU - Souza, Cleonice de
AU  - Souza Cd
FAU - Moliterno, Ricardo Alberto
AU  - Moliterno RA
FAU - Visentainer, Jeane Eliete Laguila
AU  - Visentainer JE
LA  - por
PT  - English Abstract
PT  - Journal Article
TT  - Extracao de DNA a partir de sangue humano coagulado para aplicacao nas tecnicas 
      de genotipagem de antigenos leucocitarios humanos e de receptores semelhantes a 
      imunoglobulina.
PL  - Brazil
TA  - Rev Soc Bras Med Trop
JT  - Revista da Sociedade Brasileira de Medicina Tropical
JID - 7507456
RN  - 0 (HLA Antigens)
RN  - 0 (Receptors, KIR)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - DNA/blood/*isolation & purification
MH  - Genotype
MH  - HLA Antigens/*genetics
MH  - Humans
MH  - Luminescent Measurements
MH  - Polymerase Chain Reaction/methods
MH  - Receptors, KIR/*genetics
EDAT- 2010/03/09 06:00
MHDA- 2010/05/11 06:00
CRDT- 2010/03/09 06:00
PHST- 2009/05/18 00:00 [received]
PHST- 2009/10/23 00:00 [accepted]
PHST- 2010/03/09 06:00 [entrez]
PHST- 2010/03/09 06:00 [pubmed]
PHST- 2010/05/11 06:00 [medline]
AID - S0037-86822009000600008 [pii]
AID - 10.1590/s0037-86822009000600008 [doi]
PST - ppublish
SO  - Rev Soc Bras Med Trop. 2009 Nov-Dec;42(6):651-6. doi: 
      10.1590/s0037-86822009000600008.