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利用超深度焦磷酸测序和系统进化分析鉴定三起诉讼案中的 HIV-1 传播源。

Source identification of HIV-1 transmission in three lawsuits Using Ultra-Deep pyrosequencing and phylogenetic analysis.

机构信息

Center for Infectious Disease and Cancer Research, Kaohsiung Medical University, Kaohsiung, Taiwan; Graduate Institute of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.

Center for Infectious Disease and Cancer Research, Kaohsiung Medical University, Kaohsiung, Taiwan; Basic Research Laboratory, National Cancer Institute, National Institutes of Health, Frederick, MD, USA.

出版信息

J Microbiol Immunol Infect. 2021 Aug;54(4):596-605. doi: 10.1016/j.jmii.2019.12.002. Epub 2020 Jan 27.

Abstract

BACKGROUND/PURPOSE: Intentional transmission of HIV-1 is a crime. Identifying the source of transmission between HIV-1 infected cases using phylogenetic analysis has limitations, including delayed examinations after the initiation of infection and ambiguity of phyletic relationships. This study was the first to introduce phylogenetic tree Results as forensic evidence in a trial in Taiwan.

METHODS

Three lawsuit cases from different district courts in Taiwan were chosen for this study. We identified the source of transmission between individuals in each lawsuit based on the maximum likelihood and Bayesian phylogenetic tree analyses using the HIV-1 sequences from molecular cloning and ultra-deep pyrosequencing (UDPS). Two gene regions of the HIV genome, env and gag, were involved.

RESULTS

The results of phylogenetic analysis using sequences from molecular cloning were clear and evidential enough in lawsuits 1 and 3. Due to the delayed sampling time, the result of sequences from molecular cloning in lawsuit 2 was ambiguous. Combined with the analyzed result of sequences from UDPS and epidemiological information, the source of transmission in lawsuit 2 was further identified.

CONCLUSION

Hence phylogenetic analyses cannot exclude the possibility of unsampled intermediaries, the data interpretation should be more careful and conservative, and it should not be considered as the only evidence for the source identification in a trial without epidemiological or serological information. The evaluation of the introduced UDPS method in the identification of transmission source has shown that the validity and evidential effects were still limited and need further optimization.

摘要

背景/目的:故意传播 HIV-1 是犯罪。通过系统发育分析确定 HIV-1 感染病例之间的传播源存在局限性,包括感染开始后延迟检查以及系统发育关系的模糊性。本研究首次将系统发育树结果作为法庭证据引入台湾的一项审判中。

方法

选择了来自台湾不同地区法院的三个诉讼案件进行本研究。我们根据来自分子克隆和超深度焦磷酸测序(UDPS)的 HIV-1 序列,使用最大似然和贝叶斯系统发育树分析,确定了每个诉讼中个体之间的传播源。涉及 HIV 基因组的两个基因区域,env 和 gag。

结果

在诉讼 1 和 3 中,使用分子克隆序列进行系统发育分析的结果清晰且具有足够的证据力。由于采样时间延迟,诉讼 2 中分子克隆序列的结果存在模糊性。结合 UDPS 分析结果和流行病学信息,进一步确定了诉讼 2 中的传播源。

结论

因此,系统发育分析不能排除未采样中间人的可能性,数据解释应更加谨慎和保守,并且在没有流行病学或血清学信息的情况下,不应将其视为审判中来源识别的唯一证据。引入 UDPS 方法在鉴定传播源中的有效性和证据效力仍有限,需要进一步优化。

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