Screening for barbiturates in vitreous humor by the EMIT-st serum enzyme immunoassay

In 16 medical examiner's cases, which were found to be barbiturate-positive by thin-layer chromatographic screening of the liver, blood barbiturate concentrations were determined by gas chromatography. The corresponding vitreous humor samples were screened by the enzyme multiplied immunoassay technique, the EMIT-st serum barbiturate assay. By using the recommended dilution for detecting serum barbiturates, it was possible to detect barbiturates in vitreous humor at a toxic concentration. By using one fourth the amount of diluent, the barbiturates could be detected also at a therapeutic concentration. The EMIT-st assay proved to be useful in the screening for barbiturates in vitreous humor, a material that is readily available in forensic toxicology.     

Developmental Validation of the Quantifiler® Duo DNA Quantification Kit for Simultaneous Quantification of Total Human and Human Male DNA and Detection of PCR Inhibitors in Biological Samples*

Abstract: The Quantifiler^® Duo DNA Quantification kit enables simultaneous quantification of human DNA and human male DNA as well as detection of inhibitors of PCR in a single real-time PCR well. Pooled human male genomic DNA is used to generate standard curves for both human (ribonuclease P RNA component H1) and human male (sex determining region Y) specific targets. A shift in the cycle threshold (C_T) values for the internal positive control monitors the presence of PCR inhibitors in a sample. The assay is human specific and exhibits a high dynamic range from 0.023 to 50 ng/μL. In addition, the multiplex assay can detect as little as 25 pg/μL of human male DNA in the presence of a 1000-fold excess of human female DNA. The multiplex assay provides assessment of the DNA extract and guidance for the selection of the appropriate AmpFℓSTR^® Amplification Kit to obtain interpretable short tandem repeat profiles.     

A comparison of methods used in the UK and Ireland for the extraction and detection of semen on swabs and cloth samples

The recent formation of a United Kingdom and Irish working group, the Body Fluids Forum (BFF), highlighted the need to investigate different working practices prior to any inter-laboratory comparison work and identification of best practice. Various dilutions of semen were seeded onto swabs and cloth samples for each BFF member laboratory to test using their standard techniques. The results showed that the detection of acid phosphatase on swabs is best achieved using direct testing rather than on an extract from the swab. Extraction methods for spermatozoa require a balance to be achieved between using a sufficient volume of water to ensure optimal release and minimal volume to ensure a concentrated extract. PSA tests were investigated and found to be more sensitive than Choline. DNA profiles were obtained from samples in which no spermatozoa had been detected during microscopic examination.