mRNA profiling for body fluid identification by multiplex quantitative RT-PCR.
An alternative approach to conventional protein-based body fluid identification is gene expression profiling analysis. In the present work, we report the development of sensitive and robust multiplex quantitative reverse transcriptase-PCR assays for the identification of blood, saliva, semen, and menstrual blood. Each body fluid assay comprises a triplex system that detects transcripts from two body fluid-specific genes and a housekeeping gene GAPDH. The body fluid-specific genes include erythroid delta-aminolevulinate synthase (ALAS2) and beta-spectrin (SPTB) for blood, statherin (STATH) and histatin 3 (HTN3) for saliva, protamine 1 (PRM1) and protamine 2 (PRM2) for semen, and matrix metalloproteinase 7 (MMP7) and matrix metalloproteinase 10 (MMP10) for menstrual blood. Normalization of both body fluid-specific genes to the housekeeping gene by means of appropriate cycle threshold metrics ensures the high specificity of each assay for its cognate body fluid.
Juusola
Jane
J
Graduate Program in Biomolecular Sciences, University of Central Florida, Orlando, FL, 32816-2366, USA.
Ballantyne
Jack
J
D000624:5-Aminolevulinate Synthetase / Q000235:genetics; D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001769:Blood; D002648:Child; D002675:Child, Preschool; D016172:DNA Fingerprinting; D005260:Female; D053803:Forensic Genetics; D020869:Gene Expression Profiling; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D054808:Histatins / Q000235:genetics; D006801:Humans; D007223:Infant; D008297:Male; D053506:Matrix Metalloproteinase 10 / Q000235:genetics; D020783:Matrix Metalloproteinase 7 / Q000235:genetics; D008598:Menstruation; D008875:Middle Aged; D011479:Protamines / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012463:Saliva; D012471:Salivary Proteins and Peptides / Q000235:genetics; D012661:Semen; D013049:Spectrin / Q000235:genetics
C068075:HTN3 protein, human; D054808:Histatins; D011479:Protamines; D012333:RNA, Messenger; C018149:STATH protein, human; D012471:Salivary Proteins and Peptides; C068621:protamine 2; D013049:Spectrin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D000624:5-Aminolevulinate Synthetase; C480079:ALAS2 protein, human; D053506:Matrix Metalloproteinase 10; D020783:Matrix Metalloproteinase 7