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형광 보합법을 이용한 사산아 태반조직의 염색체 수적이상 검출

Detection of Numerical Chromosome Aberrations Using Fluorescence in situ Hybridization in Stillbirth Placenta Tissue

대한보건연구 v.29 no.1 , 2003년, pp.71 - 80  
노명희, 조민호, 방원기
초록

Fluorescence in situ hybridization(FISH) is a very useful means of evaluating the chromosome aberrations in paraffin embedded tissue. Attempt was made to set up the standard for the adequate process of pretreatment and pepsin, which is required in the interphase FISH of paraffin embedded tissue. To understand the ratio of the aneuploidy in stillbirth chromosomes, the interphase FISH was carried out on 54 examples of stillbirth placentas which had been recognized as normal in the histomorphological examination at Samsung Seoul Hospital. The existing cytogenetic examinations have been unable to carry out examinations on the stillbirths according to the requirements of the cell culture. For that reason, the chromosome aberrations were evaluated employing the interphase FISH. With the interphase FISH, it is generally possible to examine chromosomes in a piece of a section of the paraffin embedded tissue used for examination of tissues. However, the paraffin block interphase FISH has technical disadvantages, judging from its signal, because its degree of color intensity is low under a fluorescent microscope. Furthermore, pretreatment and pepsin can help increase the degree of color intensity. This study used non-treatment, 2X SSC, and sodium thiocyante for the pretreatment, and found that the process of sodium thiocynante provided an appropriate result. However, there was a possibility that the signal would vanish through the process of pepsin, because it accompanied the nucleus morphological loss within the cell. The result of adjusted time for the process of pepsin, demonstrated that the best condition will be obtained after 10 minutes. As a result of the examination of X, 9, 16, 18 chromosome interphase FISH that reportedly broke out frequently in placenta tissues, of the 54 placenta tissues that had been judged as normal according to the histomorphological examination and clinical information, the following were observed from the 23 cases: chromosome X was demonstrated from 8 cases(35%), chromosome 9 from 4 cases(17%), chromosome 16 from 6 cases(26%) and chromosome 18 from 5 cases(22%).

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