Leanza Suzanne M, Burk Robert D, Rohan Thomas E
Marion Bessin Liver Research Center, Albert Einstein College of Medicine, Ullmann Building, Room 503, 1300 Morris Park Avenue, Bronx, NY 10461, United States.
Cancer Detect Prev. 2007;31(6):480-8. doi: 10.1016/j.cdp.2007.10.009.
Because of concerns regarding the quality and quantity of DNA isolated from human hair, such samples are often overlooked as a source of DNA for molecular epidemiological studies. Nevertheless, there are many potential benefits to using hair: it is easily self-collected; it does not require costly collection kits; it can be mailed for a nominal fee; and the hair specimens can be stored at room temperature. However, the amount of DNA that can be extracted from hair samples is somewhat limited. Therefore, we assessed the feasibility of using whole genome amplification (WGA) on genomic DNA extracted from archived human hairs (stored for 7 to 11 years) to increase the quantity of DNA available for genotyping analysis.
We evaluated two methods of WGA, multiple displacement amplification and the Genomeplex method. Both WGA methods were performed on each of 44 DNA samples isolated from archived human hair specimens. The resulting WGA products where then screened for the presence of three single nucleotide polymorphisms. The genotyping results were compared to genotyping data obtained from DNA isolated from mouthwash samples collected from the same individuals.
When we focused on DNA extracted from the hair root, we observed excellent agreement between the genotypes determined from both the hair (pre-WGA) samples and Genomeplex WGA when compared to their corresponding mouthwash DNA samples (kappa=0.83-0.91 and 0.79-0.92, respectively); whereas the agreement between the MDA samples and mouthwash DNA samples was poor (kappa=0.27-0.51).
Our data suggest that, when combined with Genomeplex WGA, hair specimens containing the root portion can serve as a reliable and renewable source of DNA.
由于对从人类头发中分离出的DNA的质量和数量存在担忧,此类样本在分子流行病学研究中常被视为DNA来源而被忽视。然而,使用头发有许多潜在的好处:它易于自行采集;不需要昂贵的采集试剂盒;可以以较低的邮费邮寄;并且头发样本可以在室温下保存。但是,从头发样本中提取的DNA量在一定程度上是有限的。因此,我们评估了对从存档的人类头发(保存7至11年)中提取的基因组DNA进行全基因组扩增(WGA)以增加可用于基因分型分析的DNA量的可行性。
我们评估了两种WGA方法,多重置换扩增和Genomeplex方法。这两种WGA方法分别对从存档的人类头发样本中分离出的44个DNA样本进行。然后对所得的WGA产物进行三种单核苷酸多态性的筛选。将基因分型结果与从同一受试者采集的漱口水样本中分离出的DNA获得的基因分型数据进行比较。
当我们关注从发根提取的DNA时,与相应的漱口水DNA样本相比,我们观察到从头发(WGA前)样本和Genomeplex WGA确定的基因型之间具有极好的一致性(kappa分别为0.83 - 0.91和0.79 - 0.92);而MDA样本与漱口水DNA样本之间的一致性较差(kappa为0.27 - 0.51)。
我们的数据表明,当与Genomeplex WGA结合使用时,包含根部的头发样本可以作为可靠且可再生的DNA来源。