用于单细胞性别鉴定的荧光原位杂交、引物原位标记以及传统和荧光聚合酶链反应的比较：对植入前基因诊断的适用性
Comparison of FISH PRINS, and conventional and fluorescent PCR for single-cell sexing: suitability for preimplantation genetic diagnosis.
作者信息
Findlay I, Corby N, Rutherford A, Quirke P
机构信息
Centre for Reproduction, Growth and Development, Leeds University, UK.
出版信息
J Assist Reprod Genet. 1998 May;15(5):258-65. doi: 10.1023/a:1022584225311.
PURPOSE
Although conventional polymerase chain reaction (PCR) was the first method used for sexing in preimplantation genetic diagnosis, fluorescent in situ hybridization (FISH) has become the method of choice. Recently two new techniques, primed in situ synthesis (PRINS) and fluorescent PCR, have been developed. This study compares the reliability and accuracy of these four techniques in single cells.
RESULTS
In buccal cells, fluorescent PCR and FISH had similar reliability (94 and 93%) and accuracy (97 and 96%) rates. The reliability and accuracy of PRINS (91 and 25%) and conventional PCR (79 and 89%) were lower. In human blastomeres, FISH and fluorescent PCR had similar reliability (100%, 717; 95%, 190/201) rates. Accuracy rates were 71% (517) and 99% (188/190) for FISH and fluorescent PCR, respectively, however, too few blastomeres were analyzed by FISH for meaningful comparison. However, when these data are compared with published data, the method of choice for blastomere sexing appears to be fluorescent PCR.
CONCLUSIONS
Flouroscent PCR has major implications for PGD.
目的
虽然传统聚合酶链反应（PCR）是植入前基因诊断中最早用于性别鉴定的方法，但荧光原位杂交（FISH）已成为首选方法。最近开发了两种新技术，即引物原位合成（PRINS）和荧光PCR。本研究比较了这四种技术在单细胞中的可靠性和准确性。
结果
在颊细胞中，荧光PCR和FISH的可靠性（分别为94%和93%）和准确性（分别为97%和96%）相似。PRINS（分别为91%和25%）和传统PCR（分别为79%和89%）的可靠性和准确性较低。在人卵裂球中，FISH和荧光PCR的可靠性相似（分别为100%，717个；95%，190/201）。FISH和荧光PCR的准确率分别为71%（517个）和99%（188/190），然而，通过FISH分析的卵裂球数量太少，无法进行有意义的比较。但是，当将这些数据与已发表的数据进行比较时，卵裂球性别鉴定的首选方法似乎是荧光PCR。
结论
荧光PCR对植入前基因诊断具有重要意义。
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