Use of sequence variation in three highly variable regions of the mitochondrial DNA for the discrimination of allogeneic platelets.
BACKGROUND
Human mitochondrial DNA (mtDNA) polymorphisms can be used to detect allogeneic transfused platelets. To increase the number of informative polymorphisms we investigated three hypervariable regions (HVR1, HVR2, and HVR3) within the displacement loop (D-loop) region of the mtDNA.


METHODS
mtDNA was obtained from 119 unrelated blood donors. Forward and reverse primers were designed and conditions optimized to amplify and sequence the template mtDNA by dye terminator cycle sequencing.


RESULTS
We established a sequencing protocol for all three HVRs of the mtDNA. Polymorphic sites were found in all three regions: 66 in HVR1, 44 in HVR2, and 18 in HVR3. Combining the sequence information of HVR1, -2, and -3 resulted in 105 different genotypes of which 95 were unique. We were able to discriminate between two randomly chosen individuals with a random match probability of 1.2 percent.


CONCLUSIONS
The D-loop region of mtDNA contains a wealth of informative molecular markers for chimerism and survival studies after transfusions of cellular blood components.
Warner
Jessica B
JB
Medical Biology Division, University of Groningen, the Netherlands.
Bruin
Elinda J
EJ
Hannig
Horst
H
Hellenkamp
Frank
F
Hörning
André
A
Mittmann
Karin
K
van der Steege
Gerrit
G
de Leij
Lou F M H
LF
Garritsen
Henk S P
HS
D001483:Base Sequence; D022801:Complementarity Determining Regions / Q000235:genetics*; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000097:blood / Q000235:genetics* / Q000302:isolation & purification; D014644:Genetic Variation; D006801:Humans; D017713:Platelet Transfusion; D011336:Probability; D014184:Transplantation, Homologous / Q000502:physiology
D022801:Complementarity Determining Regions; D017931:DNA Primers; D004272:DNA, Mitochondrial