Robust and efficient direct multiplex amplification method for large-scale DNA detection of blood samples on FTA cards

• Published in: 核技术（英文版） 2013-06, Vol.24 (3), p.72-80
• Main Author: JIANG Bowei XIANG Fawei ZHAO Xingchun WANG Lihua FAN Chunhai
• Format: Article
• Language: English
• Subjects: DNA损伤; DNA检测; FTA; 多重PCR; 稳健; 脱氧核糖核酸; 血液样本; 计算机断层扫描
• ISSN: 1001-8042; 2210-3147

Deoxyribonucleic acid （DNA） damage arising from radiations widely occurred along with the development of nuclear weapons and clinically wide application of computed tomography （CT） scan and nuclear medicine. All ionizing radiations （X-rays, γ-rays, alpha particles, etc.） and ultraviolet （UV） radiation lead to the DNA damage. Polymerase chain reaction （PCR） is one of the most wildly used techniques for detecting DNA damage as the amplification stops at the site of the damage. Improvements to enhance the efficiency of PCR are always required and remain a great challenge. Here we establish a multiplex PCR assay system （MPAS） that is served as a robust and efficient method for direct detection of target DNA sequences in genomic DNA. The establishment of the system is performed by adding a combination of PCR enhancers to standard PCR buffer. The performance of MPAS was demonstrated by carrying out the direct PCR amplification on 1.2 mm human blood punch using commercially available primer sets which include multiple primer pairs. The optimized PCR system resulted in high quality genotyping results without any inhibitory effect indicated and led to a full-profile success rate of 98.13%. Our studies demonstrate that the MPAS provides an efficient and robust method for obtaining sensitive, reliable and reproducible PCR results from human blood samples.