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mRNA Profiling for Body Fluid Identification by Multiplex Quantitative RT‐PCR *
Author(s) -
Juusola Jane,
Ballantyne Jack
Publication year - 2007
Publication title -
journal of forensic sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.715
H-Index - 96
eISSN - 1556-4029
pISSN - 0022-1198
DOI - 10.1111/j.1556-4029.2007.00550.x
Subject(s) - body fluid , housekeeping gene , saliva , multiplex , reference genes , semen , amniotic fluid , microbiology and biotechnology , biology , gene , real time polymerase chain reaction , gene expression , genetics , fetus , biochemistry , medicine , pathology , pregnancy
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 δ‐aminolevulinate synthase (ALAS2) and β‐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.