Simultaneous identification of chromosomes 18, X and Y in uncultured amniocytes by using multi-primed in situ labelling technique

Abstract

The aim of this study is to validate the multi-PRINS (primed in situ labelling) technique for simultaneous detection of chromosomes 18, X and Y in uncultured amniocytes for prenatal diagnosis of aneuploidy. The sites of the newly synthesized DNA sequences were showed as fluorescent signals by using immunochemistry. A multi-PRINS technique was specifically performed for simultaneous detection in the same cells of three chromosome targets, e.g. chromosomes 18, X and Y. Fluorescent signals corresponding to chromosomes 18, X and Y were showed as yellow, red and green colour spots, respectively. A multi-FISH technique using chromosome 18, X and Y probes was performed for comparison. Sixty cases were analysed using both multi-PRINS and multi-FISH. Fifty to two hundred nuclei were scored for each case for each technique. In all cases, there was no significant difference in the detection of chromosomes 18, X and Y regarding the sensitivity, the specificity and the efficiency; multi-PRINS and multi-FISH yield a similar distribution of the number of spots per nucleus. Both techniques were able to identify aneuploid cases without any ambiguity. Both multi-PRINS and multi-FISH can accurately and reliably detect aneuploidies involving chromosomes 18, X and Y in uncultured amniocytes. Finally, multi-PRINS represents a faster and more cost-effective alternative to FISH for prenatal testing of aneuploidy in uncultured amniocytes.

Rapid-prenatal diagnosis through fluorescence in situ hybridization for preventing aneuploidy related birth defects

“…These modern techniques include the aforementioned FISH method, multi-primed in situ labeling (multi-PRINS), quantitative fluorescence polymerase chain reaction (QF-PCR) and multiplex ligation-dependent probe amplification (MLPA). [1119202122] There have been few studies on the applications of molecular cytogenetics for prenatal diagnostics in India. [1223] Moreover, there have been no publications in this area following the printing of the aforementioned articles, as per the MEDLINE search till December, 2010.…”

An organism arises from every nucleus.

“…Genotype-phenotype studies have shown that, only one phenotype, sex, has perfection and binary correlation with genotype [28]. Techniques such as multiple-color primed in situ labelling (multi-PRINS), multiple-fluorescence in situ hybridization (multi-FISH) and comparative genomic hybridization (CGH) provide identification of molecularly differentiated sex chromosomes and DNA composition of sex chromosomes [33,34]. It possible to create new combinations defining chromosomal details of meiotic division and utilising separation of chromosomes and their transcription.…”