Forensic applications of laser capture microdissection: use in DNA-based parentage testing and platform validation.

AIM To report on the successful use of Laser Capture Microdissection (LCM) as a tool for isolation of human chorionic villi from admixed maternal tissue. Subsequent DNA isolation for forensic short tandem repeat (STR) analysis for parentage testing was performed in two cases of alleged sexual assault of female victims. We also performed validation of the LCM instrument platform, using archival formalin-fixed human fetal products of conception (POC), for which microdissection was utilized to separate maternal (decidua) and fetal (chorionic villus) components. METHODS To isolate DNA from placental chorionic villi admixed with maternal decidua recovered after spontaneous or therapeutic abortion, LCM was used to separate fetal from maternal cells. In contrast to the relatively crude conventional microdissection performed using a narrow pipette, needle, or scalpel blade, LCM allows cell- or tissue-specific isolation of placental chorionic villi from archival paraffin-embedded tissue sections, leaving the maternal tissue intact. RESULTS After polymerase chain reaction (PCR) amplification of villi after LCM of 9-15 STR loci, the quantity and quality of DNA yielded from fetal cells isolated by LCM was sufficient for PCR analysis and successful forensic parentage testing. The validation data obtained on two sets of formalin-fixed archival POC tissues from anonymous donors demonstrated the encouraging reproducibility of these protocols and procedures. CONCLUSION We demonstrated the reliability and utility of LCM for forensic applications when high specificity of a particular analyzed cell population or tissue is required. Care must be taken during routine pathology procedures to avoid contamination of tissues with admixture of extraneous DNA.

[1]  Cheng Li,et al.  Genome-wide loss of heterozygosity analysis from laser capture microdissected prostate cancer using single nucleotide polymorphic allele (SNP) arrays and a novel bioinformatics platform dChipSNP. , 2003, Cancer research.

[2]  A. Elkahloun,et al.  In Situ Gene Expression Analysis of Cancer Using Laser Capture Microdissection, Microarrays and Real Time Quantitative PCR , 2002, Cancer biology & therapy.

[3]  P. Mangin,et al.  Parentage determination on aborted fetal material through deoxyribonucleic acid (DNA) profiling. , 1991, Journal of forensic sciences.

[4]  A. West,et al.  Multiplex DNA short tandem repeat analysis. A useful method for determining the provenance of minute fragments of formalin-fixed, paraffin-embedded tissue. , 2003, American journal of clinical pathology.

[5]  D. Sgroi,et al.  Laser Capture Microdissection: a rising tool in genetic profiling of cancer , 2004, Expert review of molecular diagnostics.

[6]  N. Dracopoli,et al.  Current protocols in human genetics , 1994 .

[7]  L. Liotta,et al.  Laser capture microdissection. , 2006, Methods in molecular biology.

[8]  P. Wiegand,et al.  DNA investigations on fetal material from paternity cases , 2005, International Journal of Legal Medicine.

[9]  F. Bieber,et al.  Interpretation of complex forensic DNA mixtures. , 2001, Croatian medical journal.

[10]  F. Bieber,et al.  Isolation of DNA from Forensic Evidence , 2000, Current protocols in human genetics.

[11]  M. Bauer,et al.  Paternity testing after pregnancy termination using laser microdissection of chorionic villi , 2002, International Journal of Legal Medicine.

[12]  C. Strom,et al.  Prenatal paternity testing with deoxyribonucleic acid techniques. , 1996, American journal of obstetrics and gynecology.

[13]  U. Motro,et al.  Chorionic villus sampling prior to pregnancy termination, a tool for forensic paternity testing. , 1999, Journal of forensic sciences.