Proviral sequences detected by polymerase chain reaction in peripheral blood cells of horses with equine infectious anemia lentivirus

1991 ◽  
Vol 117 (1-2) ◽  
pp. 109-119 ◽  
Author(s):  
Katherine I. O'Rourke ◽  
M. L. Besola ◽  
T. C. McGuire
1998 ◽  
Vol 44 (10) ◽  
pp. 2088-2093 ◽  
Author(s):  
Michiyo Urata ◽  
Machiko Wakiyama ◽  
Masanori Iwase ◽  
Makoto Yoneda ◽  
Sachiko Kinoshita ◽  
...  

Abstract An adenine-to-guanine mutation at nucleotide position (np) 3243 in the mitochondrial tRNALeu(UUR) gene is closely associated with various clinical phenotypes of diabetes mellitus. Because the mutation creates a new restriction site for the restriction enzyme ApaI, the mutation is usually detected and quantified by ApaI cleavage of the PCR products including np 3243. The sensitivity of the conventional method is, however, 5–10% heteroplasmy. The percentage of heteroplasmy of the mutation is usually highest in the affected tissues and is much lower in peripheral blood cells, which are used most frequently for the analysis. The sensitivity of the conventional method, however, is not sufficient to detect the mutation from peripheral blood cells. Utilizing ligation-mediated polymerase chain reaction, we have developed a feasible and sensitive method to detect 0.01% heteroplasmy of the 3243 mutation in peripheral leukocytes.


Blood ◽  
1994 ◽  
Vol 83 (3) ◽  
pp. 853-859 ◽  
Author(s):  
H Ohashi ◽  
T Hotta ◽  
A Ichikawa ◽  
T Kinoshita ◽  
R Taguchi ◽  
...  

To investigate clonal compositions of hematologic cells in paroxysmal nocturnal hemoglobinuria (PNH), we analyzed peripheral blood (PB) cells of 12 female patients with PNH, by clonality analysis using X-chromosome inactivation and assessment of expression of glycophosphatidylinositol-anchored proteins (GPI-APs) by flow cytometry. Southern hybridization showed that granulocytes were monoclonal in three and polyclonal in eight patients, respectively, whereas lymphocytes were polyclonal in all nine patients examined. Expressions of CD16 and CD59 on granulocytes varied greatly in seven patients examined. Clonality analysis of granulocytes by the polymerase chain reaction showed that CD59-and CD59low+ cells were monoclonal, whereas CD59+ cells were polyclonal. It was shown that PB cells are predominantly chimeric of clonal (GPI-AP-or GPI-APlow+) and nonclonal (GPI-AP+) cells in PNH, and that degrees of chimerism differ greatly from patient to patient.


Blood ◽  
1994 ◽  
Vol 83 (3) ◽  
pp. 853-859 ◽  
Author(s):  
H Ohashi ◽  
T Hotta ◽  
A Ichikawa ◽  
T Kinoshita ◽  
R Taguchi ◽  
...  

Abstract To investigate clonal compositions of hematologic cells in paroxysmal nocturnal hemoglobinuria (PNH), we analyzed peripheral blood (PB) cells of 12 female patients with PNH, by clonality analysis using X-chromosome inactivation and assessment of expression of glycophosphatidylinositol-anchored proteins (GPI-APs) by flow cytometry. Southern hybridization showed that granulocytes were monoclonal in three and polyclonal in eight patients, respectively, whereas lymphocytes were polyclonal in all nine patients examined. Expressions of CD16 and CD59 on granulocytes varied greatly in seven patients examined. Clonality analysis of granulocytes by the polymerase chain reaction showed that CD59-and CD59low+ cells were monoclonal, whereas CD59+ cells were polyclonal. It was shown that PB cells are predominantly chimeric of clonal (GPI-AP-or GPI-APlow+) and nonclonal (GPI-AP+) cells in PNH, and that degrees of chimerism differ greatly from patient to patient.


Transfusion ◽  
2003 ◽  
Vol 33 (9) ◽  
pp. 783-784 ◽  
Author(s):  
Ian.S. Bevan ◽  
Matthew R. Walker ◽  
Richard A. Daw ◽  
Andreas Bitsch ◽  
Gregor Bein ◽  
...  

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