Detection and correction of a migration anomaly on a 310 genetic analyzer

During STR analysis on the 310 Genetic Analyzer, retarded migration of GS500ROX size standards and alleles in some samples was observed. The contribution of reagents, capillary and performance optimized polymer POP 4 to the observed anomaly was experimentally eliminated. Variation in electrophoresis temperature between 55 degrees C and 65 degrees C did not alter the rate of migration of GX500ROX size standard and sample alleles. An eroded connector for the cathode mounted on the heat plate assembly caused the abnormal migration. Hence, it is important to verify the mobility of all fragments in the size standard for each sample to avoid any erroneous allele calls by the automated data analysis software.     

Loss of heterozygosity detected in a short tandem repeat (STR) locus commonly used for human DNA identification

Short tandem repeat (STR) markers are commonly used in basic genetic research and in human identification testing. Clinically, STRs can be used to study genetic alterations in tumors. A genetic deletion common to many types of cancer is referred to as the loss of heterozygosity (LOH). Numerous examples of LOH in cancer have been described and some have been mapped to areas located in close proximity to markers employed in human identity testing. Despite this fact, LOH has rarely been observed for STR loci commonly employed in forensic testing. Recently, for medico-legal purposes, we were asked to determine whether a tissue biopsy originated from a particular individual. For a reference source we assessed two specimens, one from normal tissue and one from cancerous tissue. When both reference specimens were used to generate DNA profiles, we observed LOH at one STR locus, D13S317. As demonstrated in other cancers only the cancerous biopsy demonstrated LOH. The forensic community should be cognizant of these unusual circumstances because, as identification of human DNA continues to be used more extensively, certain instances will arise in which reference material will not be readily available. In these situations, archived specimens may be employed as a reference source. Clinical specimens such as tissue biopsies should be used with caution if they have not been confirmed to contain normal tissue.     

Isolation of nitrobenzene from biological material

Toluene is recommended as an agent for extraction of nitrobenzene from biological material. Optimal conditions for nitrobenzene isolation from cadaveric human liver are defined and the results of isolation are quantitatively evaluated.     

A deadly anti-SIDS device

The diagnosis of sudden infant death syndrome (SIDS) has been an enigma to medical examiners and coroners for decades. The recent drastic decrease in the number of SIDS cases has been associated with infants sleeping supine instead of prone. The apparent relation between sleeping position and SIDS has led to the marketing of several positioning sleep aids. We report a case of the improper use of one such device which resulted in the death of an infant.     

Postmortem diagnosis of cerebral malaria

Human cerebral malaria is a frequent encephalopathy that occurs in the endemic tropical-subtropical zones. There are a smaller number of imported cases in continental zones where the diagnosis sometimes remains difficult to establish. Fifteen days after the death of a 36-year-old male French citizen in Africa, an investigation to determine the cause of death was conducted. Histologic examination of the brain permitted the diagnosis of cerebral malaria. Because of the popularity of overseas tourism and because this disorder may appear as "sudden death," these victims may be referred to a forensic pathologist. This case demonstrates the role a forensic pathologist may play in determining the cause of death in cerebral malaria.     

Segmental analysis for cocaine and metabolites by HPLC in hair of suspected drug overdose cases

Hair samples of eight postmortem cases were analyzed in segments of 1 to 3 cm for cocaine, benzoylecgonine and cocaethylene. Samples were prepared for analysis by digestion in 0.1 M HCl and subsequent extraction with mixed-mode solid-phase extraction columns. Measurement was made by reversed-phase, narrow-bore HPLC and fluorescence detection using two laboratory-made internal standards. The concentrations were in the region of 0.29-316 ng/mg of hair for cocaine, 0.43-141 ng/mg of hair for benzoylecgonine and 0.93-1.83 ng/mg of hair for cocaethylene. All eight investigated cases had cocaine-positive segments. In six of the cases, all segments were positive, suggesting regular cocaine use and two showed in-between negative segments indicating an interruption or a change of the abuse intensity. The results showed a second, remarkable observation, i.e. enormous concentration differences (factor >150) for both cocaine and benzoylecgonine between the different subjects. Furthermore, interindividual cocaine/benzoylecgonine ratios ranged from 0.02 to 8.43. We believe these observations could in part be attributed to both some of the still existing limitations in the analytical approach(es), especially the mandatory hair washing steps, and in our still too limited knowledge of the hair incorporation processes. Nevertheless, in some cases, segmental analysis proved to be an important tool to distinguish, together with postmortem examination, deadly chronic abuse from single acute drug overdosage.     

The statistical variability of blood alcohol concentration measurements in drink-driving cases

Like many other places in the world, Hong Kong has drink-driving legislation which prohibits a driver from having in his blood alcohol exceeding a prescribed limit while in control of a motor vehicle. The accuracy of measuring this alcohol concentration is obviously of prime concern as an erroneous result can avert the administration of justice. The common practice is to deduct all errors from the measured value and compare the deducted value with the prescribed limit, so that the benefit of all errors of the measurement is given to the driver. It is therefore important for any laboratory responsible for measuring blood alcohol concentrations to identify and quantify all errors associated with the measurement. The present study examined 900 blood alcohol determinations carried out by the Hong Kong Government Laboratory (HKGL) on cases of suspected drink driving. The determinations were performed by 5 different analysts with two different sets of instruments during 1995-1997. Statistical analysis indicated that the instruments had no bearing on the random error or variability and that even though analyst was a significant factor on variability, the deviation from the mean so caused was only 0.3% and of no practical significance. When the systematic error introduced by the tolerance limits of the certified alcohol standards (purchased from the Laboratory of Government Chemists, UK) was taken into account, the total uncertainty (random plus systematic errors) of an alcohol determination at 99.5% confidence level was found to be 4%. It is recommended that laboratories engaged in blood alcohol determination should adopt similar statistical treatment of their analytical results to find out the error and to ensure that the results are independent of analyst and instrument used.     

Computer-assisted facial image identification system using a 3-D physiognomic range finder

This system consists of a 3-D physiognomic range finder and a computer-assisted facial image superimposition unit. The 3-D range finder is composed of a detector for measuring facial surface and its control computer. The detector has two sinusoidal grating projection devices and two CCD cameras. The computer-assisted facial image superimposition unit consists of a host computer including a proprietary software, a flat surface color display and a color image scanner for inputting 2-D facial images of a criminal. The 3-D facial shape and texture of a suspect is obtained by using the range finder. To make the comparison between the 3-D facial image and the 2-D facial image, the 3-D facial image is first reproduced on a display of the host computer from a MO disk and then the 2-D facial image is taken with the color image scanner and reproduced on the display. The 3-D facial image is exactly adjusted to match the orientation and size of the 2-D facial image under the fine framework mode, and then the fine framework mode of 3-D facial image is converted to the fine texture image. The shape and positional relationships of facial components between the 3-D and 2-D facial images are examined by the fade-out or wipe image mode. The distance between the selected two points and angle among the selected three points on the 3-D and 2-D facial images are automatically measured for the assessment of anthropometrical data between both images. For evaluating the fit between the anthropometrical points on the 3-D and 2-D facial images, the reciprocal point-to-point difference between both images is compared.