Identification of semen in stain by determination of the specific activity of L-tartrate-inhibitable acid phosphatase

For identification of semen in stain the specific activity of L-tartrate-inhibitable acid phosphatase (ACP) was determined. With each stain extract, both enzyme activity and protein concentration were determined, and the specific activity (enzyme activity/protein concentration) was calculated. Seminal stains showed a value of 23.8 +/- 15.2 (mean +/- SD) IU/mg protein, while vaginal fluid stains showed a value of 0.088 +/- 0.049 IU/mg protein. Stains of other body fluids did not show any L-tartrate-inhibitable ACP activity. Furthermore, only eight of 30 plant juice stains showed any levels of L-tartrate-inhibitable ACP, although all plants tested showed ACP activity. As the present method enables us to analyze forensic samples quantitatively, it seems to be useful for forensic practice.     

The acid phosphatase test two minute cut-off: An insufficient time to detect some semen stains

The ability to detect semen in sexual offence cases is a crucial first step to locating stains which may be suitable for DNA profiling. Since the development of the acid phosphatase test in the late1950s by Stuart S. Kind, the process undertaken to perform the test has gone largely unchanged. The method currently accepted by operational forensic science laboratories allows 2 min for a reaction to be obtained, and until relatively recently, this has not been challenged. In this research, samples of semen were obtained from three donors and a range of dilutions for each sample were prepared. Each dilution was subjected to acid phosphatase testing using both direct testing and the ‘press test’ method. The results showed that semen could be detected in excess of 15 min in dilutions up to 1 in 400 using the press test method and in dilutions up to 1 in 1000 using the direct method. Of further significance was the observation that using the press test method, the two minute cut-off was insufficient to detect the majority of stains and in some cases, semen stains as strong as 1 in 20 dilutions. This research provides compelling evidence for protocols currently utilised in forensic practice to be reviewed in order that forensic scientists do not overlook potential evidential material that may prove suitable for body fluid identification such as DNA STR profiling.     

Comparison of p30 and acid phosphatase levels in post-coital vaginal swabs from donor and casework studies

The application of p30 detection to casework analysis of seminal traces on vaginal swabs is reported and compared with the levels of acid phosphatase determined.A simple crossed-over immunoelectrophoresis system was used for batch identification of swab extracts using a commercially obtained anti-p30 serum.Positive p30 results were obtained in less than 20% of the casework swab extracts, but they provide conclusive proof of the presence of semen which is a substantial advantage over the quantitative determination of acid phosphatase.     

Persistence of spermatozoa and prostatic acid phosphatase in specimens from deceased individuals during varied postmortem intervals

The survival of spermatozoa and the persistence of prostatic acid phosphatase has been an area of interest for investigators of sexual assault. However, not much documentation exists concerning the examination of a deceased individual with regard to the postmortem interval and presence of such evidence. The authors reviewed cases referred to the medical examiner's office during a 10-year period. During this time, 199 cases were both autopsied and examined for sexual assault. In particular, these examinations included procurement of swabs for Papanicolaou staining of smears and for quantitation of prostatic acid phosphatase. Most of the victims were female, although a few were male. In the majority of cases, the swabs for smears and prostatic acid phosphatase were taken from oral, vaginal, and anorectal areas in females and oral and anorectal areas in males. The smears all were stained with the routine Papanicolaou stain, and intact spermatozoa and spermatozoan heads were sought. The prostatic acid phosphatase was analyzed by the microparticle enzyme immunoassay method and reported as ng/ml. A level of greater than 100 ng/ml was considered positive. The cases were analyzed with respect to postmortem interval; presence or absence of intact spermatozoa or spermatozoan heads; presence of an elevated prostatic acid phosphatase; body location of the specimen; the time of year; location of the victim; and physical injury (anogenital) of sexual assault. The authors hope that by examining the laboratory evidence of sexual assault, a correlation can be drawn between the presence or absence of such evidence and the aforementioned variables.     

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.     

Evaluation of the modified zinc test and the acid phosphatase test as preliminary screening methods in sexual assault case material.

The modified zinc test and a commercially available acid phosphatase test were compared as to their screening parameters according to the microscopical finding of spermatozoa in cases of alleged sexual assault. The study involved 65 pieces of evidence material. It was found that the modified zinc test has a higher sensitivity and higher predictive values than the acid phosphatase test. However, when both tests are combined in parallel, the sensitivity and the negative predictive value could be raised to 99%. This finding suggests that a negative result obtained from the parallel combination of the modified zinc test and the acid phosphatase test predicts very well that no spermatozoa will be found at microscopical examination. Since the latter technique is the only one to give absolute proof of the seminal origin of stains or traces, the parallel combination of the zinc test and the acid phosphatase test might be useful in sorting out these cases or materials that deserve further investigation by more elaborate techniques.     

FISH联用激光捕获显微分离技术对混合斑进行分离检验

目的建立荧光原位杂交(FISH)联用激光捕获显微分离技术(LCM)精确分离男女混合斑中精子的检验方法。方法收集健康志愿者精子和女性阴道上皮细胞制备模拟混合斑,经过预处理后用Vysis CEPX SpectrumOrangeTMY SpectrumGreenTM试剂盒进行荧光原位杂交,并用PALM激光捕获显微分离系统分离男、女性细胞,使用Identifiler试剂盒结合低体积扩增技术分别对男女成分进行STR扩增。结果荧光原位杂交后,可清晰分辨混合斑中的男女细胞。捕获20个精子细胞可以得到完整的STR分型,检出率为80%。随着精子数目增多,检出率提高而等位基因丢失率降低。30个精子检出率最高,为95%。结论激光捕获显微分离联用FISH技术可用于混合斑中男女细胞DNA的分离检验。     

The detection of soluble ABH blood group substances in semen and saliva using monoclonal blood grouping reagents

Using an enzyme-linked immunosorbent assay (ELISA), this study investigated the use of monoclonal antibodies for detecting secreted ABH blood group substances in semen and saliva. The results demonstrated that the behavior of some monoclonals were unpredictable and often failed to detect the corresponding antigen in a number of the specimens tested. The suitability of the monoclonal reagents for detecting soluble blood group antigens could not be predicted by their behavior with red cell antigens. Consequently, care must be taken in the selection of monoclonal reagents for use in the detection of secreted blood group antigens.