Development and validation of a multiplex assay system for simultaneous detection of HIV-1, HBV, and human STRs for...

Human immunodeficiency virus (hereafter, "HIV") and hepatitis B virus (hereafter, "HBV") are prevalent infectious pathogens. In many countries, criminal law applies to those who intentionally transmit or expose others to these two dangerous pathogens. Viral infection is a significant characteristic in forensic investigations, and there are few reports on the simultaneous detection of DNA viruses, RNA viruses, and human short tandem repeats (hereafter, "STRs") in forensic laboratories. We developed a novel multiplex assay system that simultaneously identifies HIV-1, HBV, and STRs via capillary electrophoresis (hereafter, "CE"). This system demonstrated high species specificity, detecting HIV-1 RNA and HBV DNA at concentrations as low as 10 IU/mL. Full STR profiles were generated using 62.5 pg of template DNA. The combined power of discrimination of the 16 STR loci met the requirements of forensic applications. The comprehensive validation results confirmed that the multiplex assay system, which can provide qualitative results for three types of targets in a single tube via a one-step amplification process, is valuable for forensic applications. This system can offer additional individual characteristics (viral infections) for forensic DNA analysis, thus improving the efficiency of forensic investigations.