NASS唾液卡在唾液核酸采集中的应用研究

摘要

ABSTRACT:ObjectiveTo investigate the applicability of NASS card in the collection of nucleic acid in saliva for DNA testing.MethodsAnti-bacterial and mildew-proof tests were respectively carried out on NASS and FTA cards. 21 samples of saliva collected individually with NASS and FTA cards were directly ampliifed with the Identiifler plus kit to measure the peak areas of STR loci, and test was adopted in pairs to compare the corresponding data from the two cards. 1000 samples of saliva collected separately by NASS and FTA cards were directly ampliifed with the Identiifler plus kit to verify their STR proifles.ResultsFor the anti-bacterial tests, the inhibitory rate of NASS card exceeded 99.9 % againstEscherichia coli, and that of the FTA card was less than 94.4 %, with the two cards showing their inhibitory rates greater than 99.9 % against either theStaphylococcus aureus orCandida albicans. On the mildew-proof test, both the NASS and FTA card completely restrained mildew occurring within 14 days. Up to 21 days, the moldy occurrence of NASS card was level 2 and that of the FTA card at level 3. After 28 days passing, the moldy occurrence of the two cards both reached at level 4. The inhibitory effects of the two cards on PCR were observed by their inverse products’ STR loci peak areas, with larger peak areas being obtained from NASS card than those from FTA card (0.010.05). For PCR batch examination, the detection rate of the NASS card was 98.3 %and that of the FTA card was 97.9 %.Conclusion The NASS card showed better performance than the FTA card on their antibacterial and mildew-proof ability as well as the counter-inhibition of PCR, demonstrating it applicable to saliva nucleic acid collection for forensic DNA detection and analysis.%目的：探讨NASS唾液卡在唾液核酸采集中的应用效果。方法对NASS唾液卡与FTA唾液卡进行抗菌能力测试和防霉能力测试；使用两种唾液卡分别采集21份人员口腔拭子，通过Identiifler Plus试剂盒进行PCR直接扩增，对STR基因座峰面积进行配对t检验；使用两种唾液卡各采集1000份人员口腔拭子，通过Identiifler Plus试剂盒复合扩增检测STR分型。结果抗菌能力测试中NASS唾液卡对大肠杆菌的抑菌率大于99.9%，FTA唾液卡对大肠杆菌的抑菌率大于94.4%，对金黄色葡萄球菌、白色念珠菌两种唾液卡抑菌率均大于99.9%；防霉能力测试中，21天时NASS唾液卡的长霉级别为2级，FTA唾液卡为3级，其他条件下二者无差别；NASS唾液卡和FTA唾液卡对PCR均存在一定程度的抑制，NASS唾液卡在D7S820、D13S317、TPOX、FGA基因座峰面积大于FTA唾液卡（0.010.05）；批量检测中，NASS唾液卡检出率为98.3%，FTA唾液卡检出率为97.9%。结论NASS唾液卡在抗菌、防霉能力以及对PCR扩增的抑制程度方面优于FTA唾液卡，适合在唾液核酸采集中应用。