Validation of the determination of Delta F508 mutations of the cystic fibrosis gene in over 11000 mouthwashes
Validation of the determination of Delta F508 mutations of the cystic fibrosis gene in over 11000 mouthwashes
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DOI：
10.1007/s004390050045
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发表时间：
1996-03-01
期刊：
影响因子：
5.3
通讯作者：
tenKate, LP
中科院分区：
文献类型：
--
作者：
deVries, HG;Collee, JM;tenKate, LP
漱口水可用作突变检测的DNA资源，并且由于收集和DNA分离简单且便宜，它们尤其可用于大量样品。为了确定失败率（口腔样本中未获得PCR产物的比例）和大量样本中颊上皮细胞突变检测的特异性，我们收集了11413名献血者的漱口水和血液样本，并测试了漱口水的Delta F508突变，该突变在荷兰囊性纤维化染色体中的估计频率为75%。只有在漱口水中发现突变或未能获得PCR产物的情况下，才对血样进行Delta F508突变检测。该测试的敏感性是在早期家庭研究中已知的75名Delta F508携带者的漱口水中确定的。这些样本在献血者漱口之间盲法提供。漱口过程的特异性和敏感性均为100%。总失败率为5.6%。这个庞大的数字主要是由于一个特定血库的口腔冲洗不足造成的。排除该血库的结果将失败率降低至1.8%。我们的研究结果还证实，对于大量样本的漱口水程序是适合突变检测，并与适当的指示，可用于社区筛查。
Mouthwashes can be used as a DNA resource for mutation detection and, because collection and DNA isolation is simple and cheap, they could in particular, be used for large numbers of samples. To determine the failure rate (the proportion of mouth samples in which no PCR product was obtained) and the specificity of buccal epithelial cell mutation detection in large numbers of samples, we collected mouthwashes and blood samples from 11413 blood donors and tested the mouthwashes for the Delta F508 mutation, which has an estimated frequency of 75% among cystic fibrosis chromosomes in The Netherlands. Blood samples were tested for the Delta F508 mutations only if the mutation was identified in the mouthwash or in the case of a failure to obtain PCR products. The sensitivity of the test was determined in mouthwashes of 75 Delta F508 carriers known from earlier family studies. These samples were offered blindly between the mouthwashes of the blood donors. Both specificity and sensitivity of the mouthwash procedure were 100%. The overall failure rate was 5.6%. This large figure was caused mainly by insufficient rinsing of the mouth in one particular blood bank. Exclusion of the results of this blood bank reduced the failure rate to 1.8%. Our results also confirm that for a large number of samples the mouthwash procedure is suitable for mutation detection and, with proper instructions, can be used in community screening.