运用二代测序技术拆分混合STR分型

目的探讨二代测序技术在混合STR分型拆分中的应用。方法对一例强制猥亵妇女案中受害人颈部的拭子和血样作DNA提取,以Precision ID GlobalFiler^TM NGS STR Panel v2试剂盒制备文库,经Ion S5测序仪测序,运用Torrent_Suite_v5.2.1软件进行数据分析后,将检测基因座的序列多态STR分型与长度多态STR分型进行比较。结果在D8S1179、D21S11、D2S441、D2S1338、D10S1248五个基因座发现存在序列特异的等位基因亚型,利用这些亚型对混合STR分型进行了成功拆分。结论二代测序技术提供的等位基因序列信息可对混合STR分型的拆分起到帮助作用。     

长度多态与序列多态STR模型法庭科学参数比较

基于毛细管电泳平台进行STR基因座分型是当前个体识别的金标准。二代测序技术支持STR序列多态分型,并有可能在法庭科学领域被广泛应用。相比长度多态性,STR测序可提供更大的信息量,但相关法庭科学参数的定量计算十分必要。本文建立了简单的STR基因座模型,分别计算了长度多态和序列多态STR模型基因座的法医遗传学参数,结果表明:对于单个STR基因座,其序列多态模型的个体识别力和非父排除率相比长度多态模型更高,说明序列多态STR遗传标记识别无关个体及排除非父的能力更强。在联用15个非连锁遗传模拟基因座进行法医DNA分析时,长度多态模型和序列多态模型的累积匹配概率分别在10-18和10-26量级;而如果要达到长度多态模型15个基因座的累积匹配概率(10-18),仅需使用10个非连锁模拟序列多态STR基因座。希冀此模型比较能为二代测序STR数据的法庭科学应用提供参考。     

国产Goldeneye~(TM) 20A试剂盒性能指标验证

目的测试国产GoldeneyeTM20A试剂盒技术性能指标,评估其法医学应用能力。方法从方法学验证、准确性、峰值均衡性、灵敏度、批次间试剂及稳定性测试、耐受性、不同检材的适应性与一致性、种属特异性、混合样本等9个方面对GoldeneyeTM20A试剂盒进行测试。结果阳性DNA样本分型正确,内标和等位基因分型标准物符合要求;等位基因间的均衡性≥83%,同一荧光标记基因座间的均衡性≥55%,不同标记物间的均衡性≥52%;0.125ng DNA阳性样本可检出全部STR基因座分型,不同批次间和反复冻融后试剂盒测试可以获得正确分型,对降解检材和混有抑制剂的样本等具有一定的耐受性,能对案件中多种检材进行分型且分型结果一致,具有一定的种属特异性和混合DNA样本检测能力。结论国产GoldeneyeTM 20A试剂盒可用于法庭科学实际检案与建库。     

基于二代测序的24个STR基因座序列多态性

目的对95名中国汉族无关个体的24个STR基因座序列多态性进行调查。方法采用Thermo Fisher公司25重早期测试试剂盒进行STR基因座复合扩增,应用HID-Ion AmpliSeq~(TM)文库试剂盒进行文库构建,使用Ion PGM~(TM)基因测序仪进行测序反应,对Ion Torrent Suite~(TM) v4.6软件显示的STR基因分型结果分析评估,使用PowerStats分析软件计算法庭科学参数,并与STR长度多态性的参数进行比较。结果 24个STR基因座共观察到252个不同重复序列的等位基因。其中,12个STR基因座具有相同长度的不同等位基因核心序列,其累计随机匹配概率为3.5×10~(-15),其累计非父排除率达0.999 998 2。序列多态与长度多态STR的累计随机匹配概率相差两个数量级。结论针对调查的中国汉族人群,本文24个序列多态STR基因座具有较好的个体识别能力,该数据为法医STR基因座序列多态性研究提供很好的参考。     

差异提取试剂盒在混合斑DNA提取中的应用

目的评估差异提取试剂盒对混合斑样本中的精子和上皮细胞DNA分离提取的有效性。方法采用差异提取试剂盒,选择性裂解精细胞和上皮细胞,结合磁珠法分别对人为控制条件下制备的模拟混合样本和案件中的混合斑检材进行精细胞DNA和上皮细胞DNA的分离提取。对所提取的DNA进行定量分析和STR分型。结果该试剂盒能从精子和上皮细胞不同比例的混合斑中提取出高纯度的精细胞和上皮细胞DNA。结论该差异提取试剂盒适用于性侵害案件中混合斑检材的DNA提取。     

法医学二代测序STR分型准确度与测序深度的关联性评估

目的利用实验数据对法医学二代测序STR分型测序深度与分型结果准确度的关联性进行评估。方法使用商业化基因组DNA制备单一来源和混合的DNA样本,以Thermo Fisher公司的25重早期测试试剂盒进行目的STR片段扩增,每种扩增产物分别使用4种不同的序列标签平行建库,并控制标记每一种序列标签的文库上机量依次占一张Ion 318芯片的1/4、1/8、1/16、1/32。经Ion PGMTM基因测序仪测序,以及Ion Torrent SuiteTM软件进行数据分析;同时对庞敬博等人发表的基于相同试剂盒和测序仪检测的95名中国汉族无关个体的6928条等位基因、影子峰和噪音序列进行测序深度统计分析,寻找测序深度与STR分型准确度的关联性。结果各基因座测序深度随文库上样量减少而呈明显下降趋势。对于单一来源样本,每张芯片上样不超过8个均一化文库可实现全部基因座的完整分型;对于1∶20比例的混合DNA,每张芯片上样不超过4个均一化文库时,未发现微量组分的等位基因丢失。人群数据测序深度统计显示,该体系基因座间存在不均衡性,有必要针对各基因座分别设定分析阈值参数。结论测序深度与法医学STR分型结果的准确性密切相关,各基因座最低测序深度与平均测序深度的比值可作为设定分析阈值的重要参考指标。本研究确定的单张芯片上样数量仅适用于本实验体系,但相关实验设计和方案可供其他实验体系开展类似工作参考。     

运用法医学二代测序技术侦破19年久冷案

基于Y染色体STR(Y-STR)多态性的男性家系排查技术帮助全国各地破获了诸多冷案积案。然而对于出现明显降解的生物检材,或因检出Y-STR基因座数量过少而无法开展有效排查。STRSeqTyperY68男性家系精细化排查试剂盒,定位于二代测序技术,利用Mi Seq FGx二代测序平台可单管实现52个单拷贝Y-STR基因座、6个二拷贝Y-STR基因座、1个三拷贝Y-STR基因座和1个性别判定基因座的分型检测,并能同时支持STR长度和/或序列多态分型,全部基因座扩增子长度在350bp以下,且其中62个不大于300bp,适用于降解检材的检测。本文报道一起长达19年未破的强奸杀人案,用传统STR检测方法仅得到24个Y-STR基因座分型,部分300bp以上的Y-STR片段未检出,但通过二代测序方法使用STRSeqTyperY68试剂盒完整得到了67个Y-STR和1个性别基因座分型,从而帮助办案单位锁定了嫌疑人所在的男性家系,为案件侦破提供了关键技术支撑。     

以CE-PCR产物为模板的STR序列多态分型方法

目的实现以荧光标记复合扩增产物为扩增模板的STR序列多态分型。方法筛选常用STR分型试剂盒基因座并设计引物,构建常染色体STR复合扩增体系。分别以毛细管电泳(CE)试剂盒GlobalFiler~(TM)、PowerPlex~?21和Identifiler~(TM) Plus的扩增产物为模板,扩增、建库测序及数据分析,评估体系分型准确性及对CE扩增产物的通用性,并与Precision ID GlobalFiler~(TM) NGS STR Panel体系进行比较。结果该复合扩增体系能够从3款CE试剂盒的扩增产物中获得所有基因座的正确分型和序列多态信息,其体系均衡性和杂合子均衡性亦良好,且均优于Precision ID体系。结论该复合扩增体系建立了CE技术与二代测序技术之间的桥梁,实现了检材的零利用,能够进一步挖掘荧光标记扩增产物的STR序列多态信息,是对现有技术的提升和有效补充。     

SiFa~(TM) 23 Plex试剂盒(提取测试版)在汉族人群中的法医学调查

目的评估SiFa~(TM) 23 Plex试剂盒(提取测试版)的性能及其所包含的STR基因座在汉族人群中的群体遗传学信息。方法应用试剂盒对1 000名汉族健康无关个体进行分型检测,检测试剂盒有效性并统计分析STR基因座的频率数据及群体遗传学参数信息。结果该试剂盒最小扩增体系可以为6.25μL,标准体系25μL中DNA含量低至125 pg可获取理想分型图谱。试剂盒中包含的21个常染色体STR基因座在1000人份中共检测到267个等位基因,均符合Hardy-Weinberg平衡。新增加的两个STR基因座D1S1656和D10S1248分别检测到15和11个等位基因,多态信息含量丰富。结论 SiFa~(TM) 23 Plex试剂盒(提取测试版)对于血液样本检测性能优越,准确性、重复性、灵敏度均可满足法医学检案需求,可以应用于法医学实际检案及DNA建库工作。     

05式警用转轮手枪弹壳接触DNA检验方法的比较

目的比较05式警用转轮手枪弹壳表面接触性DNA检验方法,为实际检验提供参考和借鉴。方法制备40例击发后手枪弹壳的模拟样本,分别用两步转移法提取弹壳表面不同部位检材,采用两种DNA提取法和两种扩增试剂盒对样本进行STR分型检验,比较评价检验结果。结果避开发射药残留区域采用两步转移法提取样本,有助于提高检出率;Chelex-100联合Microcon-100法提取模板DNA的产量最高可达1.18ng,高于Mini M48试剂盒法(0.91ng);MiniFilerTM试剂盒的等位基因检出率(23.61%)高于IdentifilerTM试剂盒(6.41%)。结论采用选择适当区域提取检材,采用Chelex-100联合Microcon-100法提取DNA,经MiniFilerTM试剂盒扩增,进行弹壳接触DNA分型的效果较好。     

Bhartṛhari and Maṇḍana on Avidyā

The concept of avidyā is one of the central categories in the Advaita of Śaṇkara and Maṇḍana. Shifting the focus from māyā, interpreted either as illusion or as the divine power, this concept brings ignorance to the forefront in describing duality and bondage. Although all Advaitins accept avidyā as a category, its scope and nature is interpreted in multiple ways. Key elements in Maṇḍana’s philosophy include the plurality of avidyā, individual selves as its substrate and the Brahman as its field (viṣaya), and the distinction in avidyā between non-apprehension and misapprehension. A closer investigation shows that Maṇḍana is directly influenced by Bhartṛhari’s linguistic non-dualism in developing the concept of avidyā. This study also compares other key constituents such as vivartta and pariṇāma that are relevant to the analysis of avidyā. As the concept of counter-image (pratibimba) emerges as a distinct stream of Advaita subsequent to Maṇḍana, this study also compares the application of pratibimba in the writings of Bhartṛhari and Maṇḍana.     

Unraveling the <Emphasis Type="Italic">Kāvyaprakāśa</Emphasis>: Jayadeva Pīyūṣavarṣa’s idiosyncratic sequence of topics in the <Emphasis Type="Italic">Candrāloka</Emphasis>

In his twelfth century alaṃkāraśāstra, the Candrāloka, Jayadeva Pīyūṣavarṣa reverses the sequence of topics found in Mammaṭa’s Kāvyapr-akāśa, an earlier and immensely popular work. With such a structural revisionism, Jayadeva asserts the autonomy of his own work and puts forth an ambitious critique of earlier approaches to literary analysis. Jayadeva investigates the technical and aesthetic components of poetry in the first part of the Candrāloka, prior to his formal semantic investigations in the latter half of the text, thus suggesting that aesthetic evaluations of poetry beneficially inform scientific investigations of language. Jayadeva’s organization of his chapters on the semantic operations, moreover, intimates that the study of suggestive and metaphoric functions of language clarifies our understanding of denotation, which is conventionally understood to be the primary and direct path of verbal designation.     

The Supreme Array Scripture: A New Interpretation of the Title “Gaṇḍavyūha-sūtra”

This article argues for a new interpretation of the Sanskrit compound gaṇḍa-vyūha as it is used in the common title of the Mahāyāna text the Gaṇḍavyūha-Sūtra.The author begins by providing a brief history of the sūtra’s appellations in Chinese and Tibetan sources. Next, the meanings of gaṇḍa (the problematic member of the compound) are explored. The author proposes that contemporary scholars have overlooked a meaning of gaṇḍa occurring in some compounds, wherein gaṇḍa can mean simply “great,” “big” or “massive.” This general sense is particularly common in the compound gaṇda-śaila (a “massive rock” or “boulder”) and is found in such texts as the Bhāgavata Purāṇa, the Harivaṃśa and the Harṣacarita. Following the discussion of Gaṇḍa, the author examines the term vyūha (“array”) as it is used in the Gaṇḍavyūha-sūtra. The article concludes with the suggestion that a more appropriate translation of the Gaṇḍavyūha-sūtra would be “The Supreme array Scripture.”     

Evaluation of six presumptive tests for blood, their specificity, sensitivity, and effect on high molecular-weight DNA

Luminol, leuchomalachite green, phenolphthalein, Hemastix, Hemident, and Bluestar are all used as presumptive tests for blood. In this study, the tests were subjected to dilute blood (from 1:10,000 to 1:10,000,000), many common household substance, and chemicals. Samples were tested for DNA to determine whether the presumptive tests damaged or destroyed DNA. The DNA loci tested were D2S1338 and D19S433. Leuchomalachite green had a sensitivity of 1:10,000, while the remaining tests were able to detect blood to a dilution of 1:100,000. Substances tested include saliva, semen, potato, tomato, tomato sauce, tomato sauce with meat, red onion, red kidney bean, horseradish, 0.1 M ascorbic acid, 5% bleach, 10% cupric sulfate, 10% ferric sulfate, and 10% nickel chloride. Of all the substances tested, not one of the household items reacted with every test; however, the chemicals did. DNA was recovered and amplified from luminol, phenolphthalein, Hemastix, and Bluestar, but not from leuchomalachite green or Hemident.     

Monks Who Have Sex: Pārājika Penance in Indian Buddhist Monasticisms

In the study of Buddhism it is commonly accepted that a monk or nun who commits a pārājika offence is permanently and irrevocably expelled from the Buddhist monastic order. This view is based primarily on readings of the Pāli Vinaya. With the exception of the Pāli Vinaya, however, all other extant Buddhist monastic law codes (Dharmaguptaka, Mahāsāṅghika, Mahīśāsaka, Sarvāstivāda and Mūlasarvāstivāda) contain detailed provisions for monks and nuns who commit pārājikas but nevertheless wish to remain within the saṅgha. These monastics are not expelled. Rather, they are granted a special status known as the śikṣādattaka. In this paper I explore the rules. concerning pārājika penance and the śikṣādattaka with specific regard to monastic celibacy. Given that five out of six extant law codes recognise this remarkable accommodation to the rule of celibacy, I argue that we must look to Vinayas other than the Pāli Vinaya if we are to arrive at a nuanced and representative view of Indian Buddhist monasticism.

Grant me chastity and continence, but not yet. Augustine of Hippo (354–430 C.E.)

     

Family breakdown and the ‘Welfare Child’ in 19th and 20th century Britain

This article traces new cycles of interest in past children as distinct from past childhood. Recent work highlighting that a conceptualisation of childhood existed even in periods with few written records closes the chapter begun by Philippe Ariès in 1960. Instead, there has been a new surge of interest in children on the edges of family life, as well as children in similarly liminal positions between the worlds of adults and children: runaways, delinquents and orphans. Several themes in the literature are identified, based on the conflicting ideas of ‘body/mind’, ‘victim/threat’, ‘needs/rights’. It is noted that researchers are using more imaginative ways of reaching the lived experience of children than the family or institutional framework, and that an increasing link is drawn between historical and modern concerns such as child abuse and the care of ‘at risk’ children.     

Vedic Language and Vaiṣṇava Theology:Madhva’s Use of Nirukta in his Ṛgbhāṣya

This article explores the way in which Madhva (1238–1317), the founder of the Dvaita Vedānta system of Hindu thought, reformulates the traditional exegetic practice of nirukta or “word derivation” to validate his pluralistic, hierarchical, and Vaiṣṇava reading of the Ṛgvedic hymns. Madhva’s Ṛgbhāṣya (RB) is conspicuous for its heavy reliance on and unique deployment of this exegetical tactic to validate several key features of his distinctive theology. These features include his belief in Viṣṇu’s unique possession of all perfect attributes (guṇaparipūrṇatva) and His related conveyability by all Vedic words (sarvaśabdavācyatva). Such an understanding of Vedic language invokes the basic nirukta presupposition that words are eternally affiliated with the meanings they convey. But it is also based onMadhva’s access to a lexicon entitled Vyāsa’s Nirukti with which his critics and perhaps even his commentators seem to be unfamiliar.While the precise status of this text is the subject of ongoing debate, Madhva’s possession of special insight into the sacred canon is established in part by his unique claim to be an avatāra of the wind god Vāyu and a direct disciple of Viṣṇu Himself in the form of Vyāsa₁. Thus, Madhva’s use of nirukta invokes his personal charisma to challenge not only conventional understandings of the hymns but traditional exegetic norms. Madhva’s provision of an alternative tradition of nirukta provoked sectarian debate throughout the Vijayanagara period over the extent to which one could innovate in established practices of reading the Veda. Articulating the Veda’s precise authority was a key feature of Brahmin debates during this period and reflects both the empire’s concern with promoting a shared religious ideology and the competition among rival Brahman sects for imperial patronage that this concern elicited. By looking at how two of Madhva’s most important commentators (the 14^th-century Jayatīrtha and the 17^th-century Rāghavendra) sought to defend his niruktis, this article will explore how notions of normative nirukta were articulated in response to Madhva’s deviations. At the same time, however, examining Madhva’s commentators’ defense of his niruktis also demonstrates the extent to which Madhva actually adhered to selected exegetic norms. This reveals that discomfort with Madhva’s particular methods for deriving words stemmed, in part, from a more general ambivalence towards this exegetical tactic whose inherent open-endedness threatened to undermine the fixity of the canon’s very substance: its language. Vyāsa’s Nirukti is one of several ”unknown sources” cited in Madhva’s commentaries whose exact status continues to be debated. Some scholars (e.g. Rao, Sharma, Siauve) maintain that these texts are part of a now lost Pāṅcarātra tradition that Madhva is attempting to preserve. This may be true for many of these citations. However, in addition to claiming to be both an avatāra of Vāyu and Viṣṇu-as-Vyāsa’s student, Madhva states in several places (e.g., VTN 42, RB 162) that the canon has suffered loss during transmission and that only Viṣṇu can reveal it in its entirety. Thus, it is possible that Madhva intends texts like Vyāsa’s Nirukti to be viewed as part of an ongoing and corrective revelation, a notion that is compatible with many Vaiṣṇava traditions (Halbfass, 1991: 4).     

Policing Outlawed Desire: ‘Homocriminality’ in Beat Spaces in Australia

Drawing on interview data of gay men who have had their behavior in public spaces scrutinised by agents of the law for signs deviance, this article explores the historical characteristics of police animosity towards such conduct in Australia. This entails examining encounters between police and gay men who pursue desire in ‘beat’ (or ‘cottage’ to the use the UK term) spaces. Exploring why these outlaw gay male subjects are so abject and troubling to the law, the discussion documents how law’s desire to regulate gay men plays out in the masquerade of ‘plain-clothes’ agent provocateur operations where police entrap gay men by mimicking gay bodily appearances, gestures and mannerisms. This article also examines how police regulation of gay desire functions as a form of violence that delimits expressions of same sex desire in public spaces. A key theme that underpins the analyses in this paper is that the policing of desire in ‘beat’ spaces helps produce qualities of illicitness and dangerousness and that this, in turn, fuels the circuit of desire at play between gay men and agents of the law.

Interrupting the Myth of the <Emphasis Type="Italic">Partage</Emphasis>: Reflections on Sovereignty and Sacrifice in the work of Nancy,Agamben and Derrida

Agamben traces the bio-political essence of modern politics to the non-sacrificial killing of Homo Sacer in Roman law. Nancy, on the other hand, links the history of Western politics to the fundamental logic of sacrifice in Western metaphysics. He nevertheless contemplates the possibility that Western societies may finally have arrived at the threshold of a non-sacrificial existence. Derrida seeks to resist the sacrificial logic of Western metaphysics and politics, but nevertheless appears to accept it as an irreducible fact of human co-existence. Unlike Nancy, he envisages no actual or actualised beyond beyond the realm of sacrificial metaphysics and politics. He thus can be said to interrupt Nancy’s ‘myth’ of a non-sacrificial partage. This article compares these three philosophical stances in the hope of throwing more light on the role of sacrifice in the law and politics of our time. Professor of Law, Rand Afrikaans University. Conversations with Ann van Sevenant, Carol Clarkson, Louise du Toit, Peter Fitzpatrick, Costas Douzinas and Adam Thurschwell gave impetus to many of the themes developed in this article. Concomitant shortcomings and inaccuracies, as always, are mine.