PATERNITY ASSESSMENT IN RHESUS MACAQUES (MACACA-MULATTA) - MULTILOCUSDNA-FINGERPRINTING AND PCR MARKER TYPING

Establishing kinship relations in primates using modern molecular gene tic techniques has enhanced the ability to scrutinize a number of fund amental biological issues. We screened 51 human short tandem repeats ( STRs) for cross-species PCR amplification in rhesus macaques (Macaca m ulatta) and identified 11 polymorphic loci with heterozygosity rates o f at least 0.6. These markers were used for paternity testing in three social groups (M, R, and S) of rhesus macaques from Cayo Santiago, Pu erto Rico. Several consecutive birth cohorts were analyzed in which ap proximately 200 males were tested for paternity against more than 100 mother/infant pairs. Despite a combined exclusion rate of more than 99 .9% in all three groups, some cases could not be solved unequivocally with the STR markers and additional testing of the MHC-associated DQB1 polymorphism. A final decision became possible through multilocus DNA fingerprinting with one or more of the oligonucleotide probes (GATA)( 4), (CA)(8), and (CAC)(5). Paternity assessment by multilocus DNA anal ysis with probe (CAC)(5) alone was found to have limitations in rhesus macaques as regards the number of potential sires which might be invo lved in a given case. Multilocus DNA fingerprinting requires large amo unts of DNA, and the ensuing autoradiographic patterns present difficu lties in comparisons across gels and even within the same gel across r emote lanes. Computer-assisted image analysis was incapable of elimina ting this problem. Therefore, a dual approach to DNA typing has been a dopted, using STR markers to reduce the number of potential sires to a level where all remaining candidates can be tested by multilocus DNA fingerprinting on a single gel, preferably in lanes adjacent to the mo ther/infant pair. (C) 1998 Wiley-Liss, Inc.