Hayney M S, Poland G A, Lipsky J J
Clinical Pharmacology Unit, Mayo Clinic and Foundation, Rochester, MN 55905, USA.
Hum Hered. 1996 Mar-Apr;46(2):108-11. doi: 10.1159/000154335.
Buccal-cell-derived DNA collected by a 'swish and spit' technique and blood-derived DNA were compared for ease of collection, participant acceptance, utility and accuracy for HLA class II DR typing by the polymerase chain reaction with sequence-specific primers (PCR-SSP). The HLA class II DR typing results determined from DNA extracted by proteinase K digestion followed by phenol/chloroform extraction and ethanol precipitation from buccal cells and blood cells were identical for all subjects we studied (n = 10). Class II typing by PCR-SSP using DNA extracted from buccal cells stored at -20, 4, 25 or 37 degrees C for 1 week was successful. The samples can be collected without medical supervision and are not affected by exposure to a variety of temperature conditions for up to 1 week. The stability of the buccal cell specimens to these extreme conditions demonstrates the utility of this 'swish and spit' technique for collecting nucleated cells for geographically dispersed large-scale population studies. Buccal-cell-derived DNA collected by a simple 'swish and spit' mouthwash technique is an excellent and practical substitute for blood-derived DNA.
采用“漱吐法”收集的颊细胞来源的DNA与血液来源的DNA在收集的简易程度、受试者接受度、实用性以及通过序列特异性引物聚合酶链反应（PCR-SSP）进行HLA II类DR分型的准确性方面进行了比较。我们研究的所有受试者（n = 10）中，通过蛋白酶K消化后经酚/氯仿提取及乙醇沉淀从颊细胞和血细胞中提取的DNA所确定的HLA II类DR分型结果均相同。使用从-20℃、4℃、25℃或37℃保存1周的颊细胞中提取的DNA通过PCR-SSP进行II类分型成功。样本无需医学监督即可收集，且在长达1周的各种温度条件下不受影响。颊细胞标本在这些极端条件下的稳定性证明了这种“漱吐法”技术在为地理分布广泛的大规模人群研究收集有核细胞方面的实用性。通过简单的“漱吐法”漱口技术收集的颊细胞来源的DNA是血液来源的DNA的一种极佳且实用的替代物。
