ABSTRACT
OBJECTIVE@#To optimize low copy number (LCN) DNA analysis methods for forensic STR genotyping.@*METHODS@#Two groups of DNA sample, extracted using either Magnetic bead method or Chelex-100 methods, were previously amplified with a Identifiler PCR Amplification kit, but no genotype was detected. The DNA samples were concentrated using either a drying method or the Microcon-100 method, then amplified using an miniFiler PCR Amplification kit and genotyped.@*RESULTS@#Among the 127 DNA samples, 47 samples, previously extracted using the Magnetic bead method, were genotyped with 36% success rate. Eighty samples, previously extracted using the Chelex-100 method, were genotyped with 30% success rate.@*CONCLUSION@#The application of sample concentration methods and miniFiler kit can improve the success rate of LCN STR analysis.
Subject(s)
Humans , Blood Stains , DNA/analysis , DNA Fingerprinting/methods , DNA Primers , Forensic Genetics/methods , Genotyping Techniques/methods , Microsatellite Repeats , Polymerase Chain Reaction/methods , Saliva/chemistry , Sensitivity and Specificity , Specimen Handling/methods , Templates, GeneticABSTRACT
OBJECTIVE@#To investigate the expression of 18S rRNA and beta-actin mRNA in bloodstain between 8 and 15 days after death and extrapolate the time of bloodstain formation.@*METHODS@#RNA in dried bloodstain at different times was extracted, then quantified for 18S rRNA and beta-actin mRNA by real-time RT-PCR. The bloodstain formation time was deduced based on the changes of the ratio of 18S rRNA to beta-actin mRNA at different time points.@*RESULTS@#The ratio of 18S rRNA to beta-actin mRNA increased gradually with time, indicating that rRNA and mRNA degraded in different rate with time. CONCLUSION; The ratio of 18S rRNA to beta-actin mRNA could be used for estimating the time of bloodstain formation in some period.
Subject(s)
Humans , Actins/metabolism , Blood Stains , DNA, Complementary/genetics , Forensic Medicine/methods , Postmortem Changes , RNA, Messenger/metabolism , RNA, Ribosomal, 18S/metabolism , Real-Time Polymerase Chain Reaction , Time FactorsABSTRACT
OBJECTIVE@#The STR genotypping of trace oral epithelial cells which are microdissected by laser capture microdissection system (LCM) is explored.@*METHODS@#The oral epithelial cells are microdissected using a low-power infrared laser by VERITAS Microdissection Instrument. STR loci of Profiler Plus are detected by multiplex PCR procesures.@*RESULTS@#DNA genotyping of 7-8 oral epithelial cells are succeeded, and DNA genotyping of 3-4 oral epithelial cells are failed.@*CONCLUSION@#It is viable in genotyping of trace oral epithelial cells by Laser Capture Microdissection as a new technology of seperating single cell.
Subject(s)
Humans , Cell Separation/methods , DNA/genetics , Epithelial Cells , Genotype , Lasers , Microdissection/methods , Mouth/cytology , Tandem Repeat SequencesABSTRACT
OBJECTIVE@#To develope an easy to use, rapid and accurate test for detecting buprenorphine based on the principle of competitive immunoassay.@*METHODS@#Monoclonal antibody against buprenorphine was conjugated with colloidal gold and dispensed on the glass fiber. The complete antigen Buprenorphine-BSA and the goat anti-mouse IgG polyclonal antibody were separately sprayed on the nitrocellulose membrane as the test line (T line) and the control line (C line). The rapid test kit was the final assembled product of test strip with the plastic cover.@*RESULTS@#A total of 100 urine samples were tested for buprenorphine by immunochromatographic and GC/ MS methods. The accuracy was 99.0%. It is found the test kit can only detect by cross reaction with other 45 kind drugs.@*CONCLUSION@#Rapid test kit can detect buprenorphine in the samples in 5 minutes. The cut-off value of the test is 100 ng/mL.