Evaluation of the effect of ozone disinfection on forensic identification of blood, saliva, and semen stains

IF 1.9 4区医学 Q2 MEDICINE, LEGAL Science & Justice Pub Date : 2023-12-22 DOI:10.1016/j.scijus.2023.12.005

Vivien Fejes , Gábor Simon , Lilla Makszin , Katalin Sipos , Viktor S. Poor

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Abstract

Good laboratory practice minimizes the biological hazard posed by potentially infectious casework samples. In certain scenarios, when the casework sample is contaminated with highly contagious pathogens, additional safety procedures such as disinfection might be advised. It was previously proven that ozone gas treatment does not hamper STR analysis, but there is no data on how the disinfection affects other steps of the forensic analysis.

In this study, we aimed to assess the interference of ozone disinfection with forensic tests used to identify biological stains.

A dilution series of blood, saliva, and semen samples were pipetted onto cotton fabric and let completely dry. Half of the samples were subjected to ozone treatment, while the rest served as controls. All the samples were tested with specific lateral flow immunochromatographic assays and for specific RNA markers with quantitative real-time PCR. Additionally, luminol test was carried out on blood spots, Phadebas® Amylase Test on saliva stains, and semen stains were examined with STK Lab kit and light microscope following Christmas Tree or Hematoxylin-Eosin staining.

Ozone treatment had no detrimental effect on the microscopic identification of sperm cells. Undiluted blood samples were detected with luminol and immunoassay, but at higher dilution, the sensitivity of the test decreased after disinfection. The same decrease in sensitivity was observed in the detection of semen stains using STK Lab kit from STK® Sperm Tracker, and in the case of the immunoassay specific for prostate-specific antigen (PSA). Ozone treatment almost completely inhibited the enzymatic activity of amylase. The sensitivity of antibody-based detection of amylase was also greatly reduced. RNA markers showed degradation but remained detectable in blood and semen samples after incubation in the presence of ozone. In saliva, the higher Ct values of the mRNA markers were close to the detection limit, even before ozone treatment.

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评估臭氧消毒对血液、唾液和精液污渍法医鉴定的影响

良好的实验室操作可最大限度地减少具有潜在传染性的个案工作样本带来的生物危害。在某些情况下，当案件样本受到高传染性病原体污染时，可能会建议采用消毒等额外的安全程序。在这项研究中，我们旨在评估臭氧消毒对用于识别生物污渍的法医检验的干扰。半数样本经过臭氧处理，其余样本作为对照。所有样本都进行了特异性侧流免疫层析检测，以及特异性 RNA 标记的实时定量 PCR 检测。此外，还对血斑进行了发光酚测试，对唾液样本进行了 Phadebas® 淀粉酶测试，并使用 STK 实验室试剂盒和光镜对精液样本进行了圣诞树染色或血红素-伊红染色。未稀释的血液样本可以用发光酚和免疫测定法检测，但稀释度越高，消毒后检测的灵敏度越低。使用 STK® 精子跟踪器的 STK 实验室试剂盒检测精液染色剂和前列腺特异性抗原（PSA）免疫测定的灵敏度也出现了同样的下降。臭氧处理几乎完全抑制了淀粉酶的酶活性。基于抗体的淀粉酶检测灵敏度也大大降低。在有臭氧存在的情况下，血液和精液样本中的 RNA 标记出现降解，但仍可检测到。在唾液中，即使在臭氧处理之前，mRNA 标记的较高 Ct 值也接近检测限。

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来源期刊

Science & Justice 医学-病理学

CiteScore

4.20

自引率

15.80%

发文量

审稿时长

81 days

期刊介绍： Science & Justice provides a forum to promote communication and publication of original articles, reviews and correspondence on subjects that spark debates within the Forensic Science Community and the criminal justice sector. The journal provides a medium whereby all aspects of applying science to legal proceedings can be debated and progressed. Science & Justice is published six times a year, and will be of interest primarily to practising forensic scientists and their colleagues in related fields. It is chiefly concerned with the publication of formal scientific papers, in keeping with its international learned status, but will not accept any article describing experimentation on animals which does not meet strict ethical standards. Promote communication and informed debate within the Forensic Science Community and the criminal justice sector. To promote the publication of learned and original research findings from all areas of the forensic sciences and by so doing to advance the profession. To promote the publication of case based material by way of case reviews. To promote the publication of conference proceedings which are of interest to the forensic science community. To provide a medium whereby all aspects of applying science to legal proceedings can be debated and progressed. To appeal to all those with an interest in the forensic sciences.