Detection of species of graft in xenotransplants using arbitrary primed polymerase chain reaction.
Because of a shortage in the availability of human organs, xenografts have been attempted in humans with cardiac, renal, and hepatic failure, despite limited success. Use of xenografts, however, is regulated under law in various countries. In xenotransplant cases related to violation of transplantation law, determination of species of the source of tissue and organ(s) becomes highly essential. Random amplified polymorphic DNA (RAPD) protocols using six sets of arbitrary short-sequenced primers have been standardized for verifying claims of porcine cardiac and renal grafts in human transplantation cases. Six arbitrary primers used were found to generate unique amplicon patterns at 36 degrees C annealing temperature. Among the selected primers, a single primer set having the sequence 5'- GGTGCGGGAA -3' is found to be the most informative in discerning porcine tissue contamination in humans. The patterns obtained were consistent for a particular genome. The grafted organs in the studied case were analyzed to be of porcine origin.
Dutta
R
R
Central Forensic Science Laboratory, BPR&D, Ministry of Home Affairs, Government of India, Calcutta, West Bengal. dtubprd@giasc101.vsnl.net.in
Kashyap
V K
VK
D000328:Adult; D000818:Animals; D016172:DNA Fingerprinting / Q000379:methods; D017931:DNA Primers; D016027:Heart Transplantation / Q000473:pathology*; D006801:Humans; D016030:Kidney Transplantation / Q000473:pathology*; D016040:Lung Transplantation; D008297:Male; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods*; D013045:Species Specificity; D013552:Swine; D014183:Transplantation, Heterologous / Q000473:pathology*; D019737:Transplants / Q000145:classification*
D017931:DNA Primers