Detection and Quantification of Male-Specific Fetal DNA in the Serum of Pregnant Cynomolgus Monkeys (Macaca fascicularis)

摘要

Becauseoftheirdevelopmentalsimilaritiestohumans,nonhumanprimatesareoftenusedasamodeltostudyfetaldevelopmentforpotentialclinicalapplicationsinhumans.ThedetectionoffetalDNAinmaternalplasmaorserumoffersasourceoffetalgeneticmaterialforprenataldiagnosis.However,nosuchdatahavebeenreportedforcynomolgusmonkeys(Macacafascicularis),animportantmodelinbiomedicalresearch.Wehavedevelopedaspecific,highlysensitivePCRsystemfordetectingandquantifyingmale-specificfetalDNAinpregnantcynomolgusmonkeys.Weusedmultiplexquantitativereal-timePCRtoanalyzecell-freeDNAinmaternalbloodserumobtainedfrom46pregnantmonkeysatgestationalweeks5,12,and22.ThepresenceofSRYgeneandDYS14Ychromosomalsequenceswasdeterminedin28monkeyswithmale-bearingpregnancies.Accordingtoconfirmationoffetalsexatbirth,theprobeandprimersfordetectingtheYchromosomalregionsateachtimepointrevealed100%specificityofthePCRtestandnofalse-positiveorfalse-negativeresults.IncreasedlevelsoftheSRY-specificsequences(mean,4706copies/mLserumDNA;range,1731to12,625)andDYS14-specificsequences(mean,54,814copies/mLserumDNA;range,4175#8211;131,250copies)weredetectedatweek22.TheSRY-andDYS14-specificprobesappeartobeaneffectivecombinationofmarkersinamultiplexPCRsystem.Toourknowledge,thisreportisthefirsttodescribethedetectionofcell-freeDNAincynomolgusmonkeys.