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 independen...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.展开更多
Subtelomeric rearrangements contribute to idiopathic mental retardation （MR）, but most children with idiopathic MR do not show any chromosome abnormalities with standard cytogenetic analysis. The primed in situ labe...Subtelomeric rearrangements contribute to idiopathic mental retardation （MR）, but most children with idiopathic MR do not show any chromosome abnormalities with standard cytogenetic analysis. The primed in situ labeling （PRINS） technique, using an oligonucleotide primer complementary to the telemetric repeat sequences （TTAGGG）, can identify chromosome telomeric abnormality （deletion） in idiopathic MR children. In this study, seventy children with idiopathic MR were enrolled and subjected to PR1NS. The results showed normal karyotype in all the children, subtelomeric rearrangements （lq del and 4q del） in 2 cases, which was confirmed by fluorescence in situ hybridization （FISH）. It was concluded that PRINS is effective for the detection of subtelomeric rearrangements and may become a routine technique for cytogenetical abnormality screening.展开更多
Cytogenetic analysis was performed for the bay scallop （Argopecten irradians Lamarck 1819） and the Japanese scallop （Patinopecten yessoensis Jay 1857 ） by primed in situ labeling （PRINS） and propidium iodide （P...Cytogenetic analysis was performed for the bay scallop （Argopecten irradians Lamarck 1819） and the Japanese scallop （Patinopecten yessoensis Jay 1857 ） by primed in situ labeling （PRINS） and propidium iodide （PI） banding techniques. The PRINS analysis revealed that major rRNA genes were clustered in two loci on the telomeric regions of the short arms on two acrocentric chromosome pairs in A. irradians and on two submetacentric pairs in P. yessoensis. The histone H3 gene sites differed in number and location between these two species. The C-band-like patterns revealed by PI staining varied considerably between these two species. A. irradians displayed terminal bands at long arms on all chromosomes, centromeric bands on some pairs and interstitial bands on five pairs. P. yessoensis exhibited only centromeric bands on all chromosomes. These results would contribute to the better understanding of karyotype evolution in A. irradians and P. yessoensis.展开更多
染色体数目异常是人类染色体疾病的重要类型,经常导致胚胎丢失、胎儿流产、婴儿死亡、先天畸形和神经发育异常等出生缺陷。文章应用引物原位标记(Primed in situ labeling,PRINS)技术快速检测人类染色体非整倍性,率先采用更新的非ddNTP...染色体数目异常是人类染色体疾病的重要类型,经常导致胚胎丢失、胎儿流产、婴儿死亡、先天畸形和神经发育异常等出生缺陷。文章应用引物原位标记(Primed in situ labeling,PRINS)技术快速检测人类染色体非整倍性,率先采用更新的非ddNTP阻断的多色PRINS技术,对人类外周血淋巴细胞和精子等多种样本进行标记;然后对不同靶标序列的标记效率及不同荧光色素的发光特点通过实验进行评估,获得关于PRINS技术的多项反应原理参数,并筛选标记顺序以获得均一稳定的标记效果,最后进行临床FISH探针与PRINS的标记比较实验。通过实验比较PRINS技术与传统FISH技术之间的标记特点与差别,评估PRINS的实际应用效果。在2.5h内标记了同一精子核内的多条染色体,单色以上标记达到99%。同时在人类外周血淋巴细胞中也得到较好的标记效果。与FISH技术相比,PRINS的这些优点使得它成为诊断染色体非整倍性变异的首选技术。展开更多
文摘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.
文摘Subtelomeric rearrangements contribute to idiopathic mental retardation （MR）, but most children with idiopathic MR do not show any chromosome abnormalities with standard cytogenetic analysis. The primed in situ labeling （PRINS） technique, using an oligonucleotide primer complementary to the telemetric repeat sequences （TTAGGG）, can identify chromosome telomeric abnormality （deletion） in idiopathic MR children. In this study, seventy children with idiopathic MR were enrolled and subjected to PR1NS. The results showed normal karyotype in all the children, subtelomeric rearrangements （lq del and 4q del） in 2 cases, which was confirmed by fluorescence in situ hybridization （FISH）. It was concluded that PRINS is effective for the detection of subtelomeric rearrangements and may become a routine technique for cytogenetical abnormality screening.
文摘Cytogenetic analysis was performed for the bay scallop （Argopecten irradians Lamarck 1819） and the Japanese scallop （Patinopecten yessoensis Jay 1857 ） by primed in situ labeling （PRINS） and propidium iodide （PI） banding techniques. The PRINS analysis revealed that major rRNA genes were clustered in two loci on the telomeric regions of the short arms on two acrocentric chromosome pairs in A. irradians and on two submetacentric pairs in P. yessoensis. The histone H3 gene sites differed in number and location between these two species. The C-band-like patterns revealed by PI staining varied considerably between these two species. A. irradians displayed terminal bands at long arms on all chromosomes, centromeric bands on some pairs and interstitial bands on five pairs. P. yessoensis exhibited only centromeric bands on all chromosomes. These results would contribute to the better understanding of karyotype evolution in A. irradians and P. yessoensis.
文摘染色体数目异常是人类染色体疾病的重要类型,经常导致胚胎丢失、胎儿流产、婴儿死亡、先天畸形和神经发育异常等出生缺陷。文章应用引物原位标记(Primed in situ labeling,PRINS)技术快速检测人类染色体非整倍性,率先采用更新的非ddNTP阻断的多色PRINS技术,对人类外周血淋巴细胞和精子等多种样本进行标记;然后对不同靶标序列的标记效率及不同荧光色素的发光特点通过实验进行评估,获得关于PRINS技术的多项反应原理参数,并筛选标记顺序以获得均一稳定的标记效果,最后进行临床FISH探针与PRINS的标记比较实验。通过实验比较PRINS技术与传统FISH技术之间的标记特点与差别,评估PRINS的实际应用效果。在2.5h内标记了同一精子核内的多条染色体,单色以上标记达到99%。同时在人类外周血淋巴细胞中也得到较好的标记效果。与FISH技术相比,PRINS的这些优点使得它成为诊断染色体非整倍性变异的首选技术。
