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CLC number: Q987;R394.2

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Received: 2002-06-12

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Journal of Zhejiang University SCIENCE A 2002 Vol.3 No.5 P.617~621

http://doi.org/10.1631/jzus.2002.0617


Rapid detection of chromosome 18 aneuploidies in amniocytes by using primed in situ labeling (PRINS) technique


Author(s):  YANG Jian-bin, ZHENG Shu

Affiliation(s):  Cancer Institute, Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310009, China

Corresponding email(s):   bqyjb@mail.hz.zj.cn, zhengshu@mail.hz.zj.cn

Key Words:  Primed in situ labeling (PRINS), Prenatal diagnosis, Chromosome 18


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YANG Jian-bin, ZHENG Shu. Rapid detection of chromosome 18 aneuploidies in amniocytes by using primed in situ labeling (PRINS) technique[J]. Journal of Zhejiang University Science A, 2002, 3(5): 617~621.

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Abstract: 
This paper presents a feasible method for rapid detection of the interphase nuclei of uncultured amniocytes for chromosomes 18 by using our modified primed in situ labeling (PRINS) technique. A total of 262 independent, uncultured amniotic fluid samples were analysed in a blind fashion before the karyotype was available. In addition, 62 samples were examined by fluorescence in situ hybridization (FISH) for comparison. In more than 95% of the samples PRINS reactions with primer 18cen were successfully induced. Two samples were properly identified and correctly scored as trisomic 18. PRINS reaction could be performed automatically in less than one hour with a programmable thermocycler. Our studies showed that the PRINS technique is simple, rapid and cost-effective. It is as sensitive and specific as FISH; can enhance the accuracy of standard cytogenetic analysis; and allows identification of chromosomes 18 aneuploidies in uncultured amniocytes in significantly less time.

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Reference

[1] Coullin, P., Roy, L., Pellestor, F., Candelier, J., Bed-Hom, B., Guillier-Gencik, Z., Bernheim, A., 2002. PRINS, the other in situ DNA labeling method useful in cellular biology. Am. J. Med. Genet., 107:127-135.

[2] Eiben, B., Trawicki, W., Hammans, W., Goebel, R., Epplen, J.T., 1998. A prospective comparative study on fluorescence in situ hybridization (FISH) of uncultured amniocytes and standard karyotype analysis. Prenat. Diagn.,18: 901-906.

[3] Gosden, J., Lawson, D., 1995. Instant PRINS: a rapid method for chromosome identification by detecting repeated sequences in situ. Cytogenet. Cell Genet., 68: 57-60.

[4] Hindkjær, J., Koch, J., Terkelsen, C., Brandt, C.,Kφlvraa, S., Bolund, L., 1994. Fast, sensitive multicolor detection of nucleic acids by Primed IN Situ labeling (PRINS). Cytogenet. Cell Genet., 66: 152-154.

[5] Koch, J., Kφlvraa, S., Petersen, K., Gregersen, N., Bolund, L., 1989. Oligonucleotide-priming methods for the chromosome-specific labeling of alpha-satellite DNA in situ. Chromosoma 98: 259-265.

[6] Koch, J., Hindkjær, J., Kφlvraa, S., Bolund, L.,1995. Construction of a panel of chromosome-specific oligonucleotide probes (PRINS-primers) useful for the identification of individual human chromosomes in situ. Cytogenet. Cell Genet., 71: 142-147.

[7] Whiteman, D. A. H., Klinger, K., 1991. Efficiency of rapid in situ hybridization methods for prenatal diagnosis of chromosome abnormalities causing birth defects. Am. J. Hum Genet., 49 suppl: A1279.

[8] Wilkens, L., Tchinda, J., Komminoth, P., Werner, M., 1997. Single- and double-color oligonucleotide primed in situ labeling (PRINS): applications in pathology. Histochem. Cell Biol., 108: 439-446.

[9] Velagaleti, G. V. N., Tharapel, S. A., Tharapel, A. T., 1999. Validation of Primed In Situ labeling (PRINS) for interphase analysis: comparative studies with conventional fluorescence in situ hybridization and chromosome analyses. Cancer Genet. Cytogenet., 108: 100-106.

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