HAN Jun-ping, LI Yang, MA Yuan, SHANG Lei, LI Cai-xia, SUN Jing
Life Science Research. 2017, 21(5): 442-449.
Abstract: To date, multiplex PCR with fluorescently labeled primers has been used as an essential method for the amplification of short tandem repeats (STRs) in forensic DNA testing: nevertheless, the multiplex PCR is commonly identified as the bottleneck in the process of traditional STR workflow, which includes extraction, quantitation, amplification, separation and detection. It often takes up to three hours to complete 28~30 cycles of PCR. In order to ensure that all of the targets (typically 100~400 bp) can be effectively amplified in a balanced manner, long PCR cycling times are required by the use of thermal cyclers and heat stable polymerases. However, with the advent of modified polymerases and faster thermal cyclers, the time required for the PCR is no longer on the order of three hours, but as little as a few minutes. The application of rapid PCR, direct PCR, rapid PCR on chip and additional rapid PCR protocols in the field of DNA rapid testing, especially the progress of forensic DNA rapid detection are summarized here. Besides, it also summarizes the comparison of rapid PCR protocols for STR typing. So far, shortening the amplification time is the mainstream direction of forensic DNA technology research. In the future, the microfluidic chip-based fully automatic, portable and integrated DNA analysis system will extend the forensic DNA testing from the laboratory to crime scene, even to the daily life, and achieve a rapid real-time detection.
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