Mohsen Tabasi
Legal Medicine Organization, Iran
Title: Novel Multiplex Real-Time PCR Assay for the Microbial Monitoring of Tissues from Cadaver Donors for Transplantation
Biography
Biography: Mohsen Tabasi
Abstract
The spread of microbial agents by a cadaver donor organ can result not only in loss of the allograft but also causes of death among some recipients. Despite the shortage of cadaver organ donors, each donor must be assessed thoroughly for the potential transmission of infectious disease, because the consequences of the organ donor events can have a remarkable effect on the transplant outcome.
Testing of blood specimens from deceased donors who may donate tissue has traditionally been carried out with immunoassays to identify microbial antibodies or antigens. Usually, ELISA test is the first screening test widely used for monitoring of microbial contamination in donated tissues (Cornea, femur, tendon, heart and …) in forensic medicine organization and it present extensive information about the source of infection. Serological screening for HIV, HBs, HBc, HCV and HTLV is routinely performed after death on cadaveric serum samples yet they are not validated for this aim and may produce a considerable number of false positives. On the other hand, a large percentage of cadaveric samples are hemolyzed due to biological processes that occur immediately post-mortem and this phenomenon can interfere with the ELISA assay. Moreover, in ELISA assay infections may be missed due to early infections and low viral load that cause false-negative test results.
In this investigation, we focused on usefulness of a novel multiplex real-time PCR assay for the microbial monitoring of tissues from cadaver donors for transplantation. Recent developments in multiplex PCR make it possible to rapidly identify, genotype and quantify multiple DNA targets simultaneously in a single reaction. Multiplex qPCR is particularly well suited to applications such as gene expression analysis, SNP genotyping, copy number variation (CNV), genetically modified organism (GMO) detection, pathogen detection, and monitoring the efficacy of drug treatment. The advantages of multiplex qPCR include increased through-put, reduced reaction cost, and conservation of limited sample material. Multiplex qPCR often requires considerable optimization and individual assays must be validated prior to multiplexing. Careful assay design is critical to avoid primer complementarity and to ensure efficient amplification of each target amplicon.