Rapid human chromosome aberration analysis using fluorescence in situ hybridization.
We have used in situ hybridization of repeat-sequence DNA probes, specific to the paracentromric locus 1q12 and the telomeric locus 1p36, to fluorescently stain regions that flank human chromosome 1p. This procedure was used for fast detection of structural aberrations involving human chromosome 1p in two separate experiments. In one, human lymphocytes were irradiated with 0, 0.8, 1.6, 2.4 and 3.2 Gy of 137Cs gamma-rays. In the other, human lymphocytes were irradiated with 0, 0.09, 0.18, 2.0, 3.1 and 4.1 Gy of 60Co gamma-rays. The frequencies (per cell) of translocations and dicentrics with one breakpoint in 1p and one elsewhere in the genome were determined for cells irradiated at each dose point. These frequencies both increased with dose, D, in a linear-quadratic manner. The delta, alpha, and beta coefficients resulting from a fit of the equation f(D)=delta + alphaD + betaD2 to the translocation frequency dose-response data were 0.0025, 0.0027 and 0.0037 for 137Cs gamma-rays, and 0.0010, 0.0041, and 0.0057 for 60Co gamma-rays. The delta, alpha, and beta coefficients resulting from a fit to the dicentric frequency dose-response data were 0.0005, 0.0010 and 0.0028 for 137Cs gamma-rays and 0.0001, 0.0002 and 0.0035, for 60Co gamma-rays. Approximately 32,000 metaphase spreads were scored in this study. The average analysis rate was over two metaphase spreads per minute. However, an experienced analyst was able to find and score one metaphase spread every 10s. The importance of this new cytogenetic analysis technique for biological dosimetry and in vivo risk assessment is discussed.
Lucas
J N
JN
Biomedical Sciences Division, Lawrence Livermore National Laboratory, University of California, Livermore 94550.
Tenjin
T
T
Straume
T
T
Pinkel
D
D
Moore
D
D
Litt
M
M
Gray
J W
JW
D002588:Cesium Radioisotopes; D002869:Chromosome Aberrations; D002878:Chromosomes, Human, Pair 1 / Q000528:radiation effects*; D003037:Cobalt Radioisotopes; D005453:Fluorescence; D005720:Gamma Rays; D006801:Humans; D008214:Lymphocytes / Q000528:radiation effects*; D009693:Nucleic Acid Hybridization; D013997:Time Factors; D014178:Translocation, Genetic
D002588:Cesium Radioisotopes; D003037:Cobalt Radioisotopes