DNA analysis

DNA 分析
  • 文章类型: Journal Article
    探讨基于纳米粒子(NP)的免疫传感器在口腔癌领域的诊断和治疗潜力。PubMed,Embase,Scopus,WebofScience,和谷歌学者数据库被探索在口腔癌中的NP应用。对所有文章进行了数据提取和质量评估。在147篇文章中,17篇被纳入这篇综述。大多数研究表明,使用基于金NP的酶联免疫吸附测定,唾液分析的灵敏度和特异性提高。改善早期识别。此外,新的治疗方法,利用基于NP的免疫传感器,展示了靶向药物递送,化学-光热耦合疗法,和基因沉默。成像方法使得区分恶性和健康状态成为可能,如表面增强拉曼散射和光学相干层析成像。这些综述的发现强调了基于NP的免疫传感器在解决与诊断和治疗口腔癌相关的困难方面的转化潜力。然而,为了在临床实践中准确解释和应用基于NP的解决方案,必须彻底意识到所涉及的复杂性,本综述的综合数据支持在改善口腔癌管理的持续努力中继续研究和改进基于NP的疗法.
    To investigate the diagnostic and therapeutic potential of nanoparticle (NP)-based immunosensors in the field of oral cancer. PubMed, Embase, Scopus, Web of Science, and Google Scholar databases were explored for NP applications in oral cancer. Data extraction in terms and quality assessment of all the articles were done. Out of 147, 17 articles were included in this review. A majority of the studies showed improved sensitivity and specificity for saliva analysis using an enzyme-linked immunosorbent assay based on gold NPs, improving early identification. Additionally, novel therapeutic approaches, utilising NP-based immunosensors, demonstrated targeted drug delivery, coupled chemo-photothermal therapy, and gene silencing. Imaging methods have made it possible to distinguish between malignant and healthy states, such as surface-enhanced Raman scattering and optical coherence tomography. The reviews\' findings highlight the transformational potential of NP-based immunosensors in addressing the difficulties associated with diagnosing and treating oral cancer. However, for an accurate interpretation and application of NP-based solutions in clinical practise, it is essential to be thoroughly aware of the intricacies involved, and the synthesised data in this review support the continued investigation and improvement of NP-based therapies in the ongoing effort to improve the management of oral cancer.
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  • 文章类型: Journal Article
    兴奋剂包括使用或试图使用任何违禁方法或物质以及替代样品。因此,建议使用短串联重复(STR)分析来确定兴奋剂对照样本是否来自同一运动员.然而,已经认识到,使用可忽略量的DNA样品可能难以获得完整的STR分析。线粒体DNA(mtDNA)的特征在于其稳定性和高细胞拷贝数。因此,尿液中的mtDNA检测预计将用于分析无法使用STR分析进行分析的样品。这项研究的目的是通过进行敏感性来比较mtDNA测试与STR分析,一致性(全血,干血斑,和尿液),和案例类型研究。在敏感性研究中,与STR分析相比,mtDNA检测显示出更高的灵敏度。一致性研究表明,所有样品均与mtDNA序列和STR谱一致。在用于STR分析的一些尿液样品中发生等位基因脱落。病例类型样本研究表明,mtDNA测试可用于获得所有测试样本的DNA图谱,包括血,干血斑,尿液,针头上残留的血液,还有血迹.总之,mtDNA测试对于分析高度降解的DNA样本很有价值,比如尿样,与STR分析相比。初始测试程序应进行尿液测试,因为mtDNA是母体遗传的.在DNA匹配对运动员有害的情况下,可能需要额外的血液STR分析。
    Doping offenses involve the use or attempted use of any prohibited method or substance as well as substituting samples. Consequently, it has been recommended that short tandem repeat (STR) analysis be used to determine if the doping control samples are from the same athlete. However, it has been recognized that it may be difficult to obtain full STR analysis using negligible amounts of DNA samples. Mitochondrial DNA (mtDNA) is characterized by its stability and high cellular copy number. Therefore, mtDNA testing in urine is expected to be used to analyze samples that cannot be analyzed using STR analysis. The objective of this study was to compare mtDNA testing with STR analysis by conducting sensitivity, concordance (whole blood, dried blood spot, and urine), and case-type studies. In sensitivity studies, mtDNA testing exhibited greater sensitivity compared with STR analysis. Concordance studies indicated that all samples were consistent with the mtDNA sequences and STR profiles. Allelic dropout occurred in some urine samples that were examined for STR analysis. Case-type sample studies demonstrated that mtDNA testing could be used to obtain DNA profiles of all the samples tested, including blood, dried blood spots, urine, blood residues on needles, and blood stains. In conclusion, mtDNA testing is valuable for analyzing highly degraded DNA samples, such as urine samples, compared with STR analysis. Urine testing should be performed for the initial testing procedure, because mtDNA is inherited maternally. In situations where the DNA match is detrimental to the athlete, additional blood STR analysis may be required.
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  • 文章类型: Journal Article
    本章最初写于2011年。这个想法是在流式细胞术变得广泛使用之前和之后提供一些细胞周期分析的历史;为单一参数DNA内容分析的教育材料提供参考,以方法论的方式介绍和讨论多参数细胞周期分析,以一种休闲的风格,讨论过去40年来我们所思考的工作的各个方面,做一些实验,但没有发表。感觉有一个线性进展,从计数细胞的生长曲线开始,通过放射自显影来计数标记的有丝分裂细胞,DNA含量分析,通过免疫荧光和DNA含量分析定义的细胞周期状态,提取细胞周期表达谱,最后是概率状态建模,这应该是分析细胞计数细胞周期数据的“正确”方法。这就是本章的意义所在。2023年,我们更新了它,但是令人兴奋的是,光谱和质量细胞计数带来的广阔方面仍然年轻和发展,因此没有经过审查,reviewed,以成熟的形式呈现。
    This chapter was originally written in 2011. The idea was to give some history of cell cycle analysis before and after flow cytometry became widely accessible; provide references to educational material for single parameter DNA content analysis, introduce and discuss multiparameter cell cycle analysis in a methodological style, and in a casual style, discuss aspects of the work over the last 40years that we have given thought, performing some experiments, but didn\'t publish. It feels like there is a linear progression that moves from counting cells for growth curves, to counting labeled mitotic cells by autoradiography, to DNA content analysis, to cell cycle states defined by immunofluorescence plus DNA content analysis, to extraction of cell cycle expression profiles, and finally to probability state modeling, which should be the \"right\" way to analyze cytometric cell cycle data. This is the sense of this chapter. In 2023, we have updated it, but the exciting, expansive aspects brought about by spectral and mass cytometry are still young and developing, and thus have not been vetted, reviewed, and presented in mature form.
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  • 文章类型: Journal Article
    背景:流产和胎儿死亡在全前脑畸形的胎儿中很常见,所以基因检查通常必须在验尸环境中进行。在这些情况下使用培养的成纤维细胞进行常规核型分析的效率由于频繁的培养失败而受到限制。在目前的研究中,全前脑的存档病例,要求进行验尸基因评估并且有足够的冷冻材料,使用定量荧光聚合酶链反应(QF-PCR)技术进行重新评估。
    方法:测试染色体13、15、16、18、21、22、X、用QF-PCR技术对7例全前脑畸形的存档冷冻绒毛中分离的DNA进行了Y检测。
    结果:QF-PCR在所有7例病例中均成功。两例13三体,两例三倍体,发现一例18三体综合征,意味着71%的诊断率。QF-PCR的成功率(100%,7/7)优于常规核型分析(43%,3/7)。
    结论:使用QF-PCR技术进行快速非整倍体检测是一种简单的,可靠,具有时间和成本效益的方法足以在死后的大多数全前脑病例中完成病因调查。
    BACKGROUND: Abortion and fetal death are common in fetuses with holoprosencephaly, so genetic examinations often have to be made in a post-mortem setting. The efficiency of the conventional karyotyping using cultured fibroblasts in these situations is limited due to frequent culture failure. In the current study, archived cases of holoprosencephaly, where post-mortem genetic evaluation was requested and sufficient frozen material was available, were reevaluated using the quantitative fluorescence polymerase chain reaction (QF-PCR) technique.
    METHODS: Testing for aneuploidies of chromosomes 13, 15, 16, 18, 21, 22, X, and Y with the QF-PCR technique was carried out on DNA isolated from archived frozen chorionic villi in seven cases of holoprosencephaly.
    RESULTS: QF-PCR was successful in all seven cases. Two cases of trisomy 13, two cases of triploidy, and one case of trisomy 18 was found meaning a 71% diagnostic yield. The success rate of QF-PCR (100%, 7/7) was superior compared to conventional karyotyping (43%, 3/7).
    CONCLUSIONS: Rapid aneuploidy testing using the QF-PCR technique is a simple, reliable, time- and cost-effective method sufficient to conclude the etiologic investigation in the majority of holoprosencephaly cases post-mortem.
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  • 文章类型: Journal Article
    随着大规模平行测序在法医遗传学中的实施,对序列数据的理解必须伴随着这些进步,也就是说,准确的数据建模,以进行适当的统计分析。等位基因脱落,遗传数据中常见的随机效应,通常在STR结果的统计分析中建模。这项概念验证研究对4ng至7.82pg的标准样品的几个系列稀释液进行了测序,以使用ForenSeq™DNASignaturePrepKit评估一组常染色体STR的等位基因脱落趋势,IlluminaMiSeqFGx上的底漆组A。评估的参数包括基因座,profile,并运行特定信息。大多数等位基因脱落发生在小于31.25pg的DNA浓度中。统计结果表明需要基于STR描述符的特定位点建模,比如简单和复合重复模式。在得分等位基因的平均读段计数和基因座处的等位基因缺失之间没有发现相关性。观察到样品中等位基因脱落量和DNA起始量之间的统计相关性,样本的平均读取计数,和在流动池上生成的总读取计数。这项研究支持在测序的STR上使用片段长度分析中使用常见的等位基因缺失因子,同时包括其他基因座,样品,并运行特定信息。结果表明,在开发测序的常染色体STR的等位基因脱落模型的可能性时,可以考虑多种因素,包括基因座特异性分析,配置文件的总读取计数,和在流动池上测序的总读取计数。
    As massively parallel sequencing is implemented in forensic genetics, an understanding of sequence data must accompany these advancements, that is, accurate modeling of data for proper statistical analysis. Allelic drop-out, a common stochastic effect seen in genetic data, is often modeled in statistical analysis of STR results. This proof-of-concept study sequenced several serial dilutions of a standard sample ranging from 4 ng to 7.82 pg to evaluate allelic drop-out trends on a select panel of autosomal STRs using the ForenSeq™ DNA Signature Prep Kit, Primer Set A on the Illumina MiSeq FGx. Parameters assessed included locus, profile, and run specific information. A majority of the allelic drop-out occurred in DNA concentrations less than 31.25 pg. Statistical results indicated a need for locus-specific modeling based on STR descriptors, like simple versus compound repeat patterns. No correlation was seen between average read count of scored alleles and allelic drop-out at a locus. A statistical correlation was observed between the amount of allelic drop-out and the starting amount of DNA in a sample, average read count of a sample, and total read count generated on a flow cell. This study supports using common allelic drop-out factors used in fragment length analysis on sequenced STRs while including additional locus, sample, and run specific information. Results demonstrate multiple factors that can be considered when developing probability of allelic drop-out models for sequenced autosomal STRs including locus-specific analysis, total read count of a profile, and total read count sequenced on a flow cell.
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  • 文章类型: Journal Article
    自1960年代以来,同源输血(HBT)被用于耐力运动中的兴奋剂。血液来自兼容的捐赠者,也就是说,ABO和恒河猴血型一致的人.尽管1985年被国际奥委会禁止,但直到2003年才有检测方法可用。然后出现了使用红细胞(RBC)次要血型抗原的想法,这些抗原构成了某人的RBC的“身份”卡,以检测第二个RBC群体的存在。在将分离的RBC与针对特定次要血型抗原的8至12种一级抗体孵育后,对掺杂对照样品进行了验证的方法使用流式细胞术。荧光直方图中的主峰和次峰揭示了RBC的双群体的存在。估计灵敏度足以在几周内检测HBT。尽管该方法的复杂性和成本,在2004年申请之后,发现了几例HBT病例,但多年来病例数量迅速下降。在2010年代,开发和评估了其他检测HBT的方法:使用运动员生物护照方法的间接检测,几年后,法医DNA分析确定HBT后血液样本中存在两种不同的DNA。尽管后者具有很高的特异性,这种敏感性最近在体内受到质疑。如今,流式细胞术方法仍然是HBT检测的首选方法,最近的研究有助于简化该方法并提高其特异性和敏感性。
    Homologous blood transfusion (HBT) is used for doping in endurance sports since the 1960s. The blood comes from a compatible donor, that is, someone with a compatible ABO and rhesus blood group. Despite been prohibited by the IOC in 1985, no detection method was available until 2003. Then came the idea to use red blood cells (RBC) minor blood groups antigens that constitute an \"identity\" card of someone\'s RBC to detect the presence of a second RBC population. The method validated for doping control samples uses flow cytometry after incubation of isolated RBC with eight to 12 primary antibodies against specific minor blood groups antigens. The presence of double populations of RBC is revealed by a major and a minor peak in a fluorescence histogram. The sensitivity was estimated sufficient to detect HBT for a few weeks. Despite the complexity and cost of the method, right after its application in 2004, several cases of HBT were identified but the number of cases dropped rapidly over the years. In the 2010s, other ways to detect HBT were developed and evaluated: indirect detection using the Athlete Biological Passport approach, and a few years later forensic DNA analysis to establish the presence of two different DNA in a blood sample after HBT. Despite the high specificity of the latter, the sensitivity was recently questioned in vivo. Nowadays, the flow cytometry method remains the method of choice for HBT detection and recent investigations helped to simplify the method and increase its specificity and sensitivity.
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  • 文章类型: Journal Article
    在2011年福岛第一核电站(FDNPP)事故之后,福岛撤离区(FEZ)内的野猪(Susscrofa)种群的规模和分布大大增加。不断增长的人口和他们从FEZ的潜在扩散,它们暴露在高水平的放射性核素中,进入周围景观强调需要更好地了解公猪运动模式,以便制定管理公猪肉运输限制的政策并制定管理策略。在这项研究中,我们使用线粒体DNA控制区的序列数据和使用348只公猪样本的MIG-seq分析来量化福岛县及其周边地区公猪的遗传种群结构,以阐明公猪的扩散模式。在公猪样本中,检测到7个亚洲单倍型和1个欧洲单倍型。欧洲单倍型起源于事故发生后疏散区的家猪和本地公猪之间的杂交,并在广泛的地理区域的15个样本中检测到。我们的MIG-seq分析显示,居住在东部地区(包括FEZ)的公猪的遗传结构显着不同。即,东进化枝)和西方(即,福岛县的西进化枝)地区。此外,我们使用Nihonmatsu市的MIG-seq遗传数据研究了公猪扩散与公猪肌肉中铯(Cs)-137活性浓度之间的关系,位于福岛中北部地区。高Cs-137活性浓度,超过1000Bq/kg,在公猪肌肉中,在局部区域内属于东部进化枝的可能性很高。因此,我们的结果提供了FEZ公猪个体或后代扩散的空间尺度的证据。这项研究的结果还表明,个体在具有不同Cs-137污染水平的区域之间的分散是导致局部区域内公猪肌肉中Cs-137活性浓度变化的最大因素之一。
    After the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident in 2011, the wild boar (Sus scrofa) population within the Fukushima Evacuation Zone (FEZ) increased substantially in size and distribution. This growing population and their potential dispersal from the FEZ, where they are exposed to high levels of radionuclides, into the surrounding landscape underscores the need to better understand boar movement patterns in order to establish policies for managing shipping restrictions for boar meat and develop management strategies. In this study, we quantified the genetic population structure of boar in and around Fukushima prefecture using sequence data of the mitochondrial DNA control region and MIG-seq analysis using 348 boar samples to clarify boar dispersal patterns. Among boar samples, seven Asian haplotypes and one European haplotype were detected. The European haplotype originated from hybridization between domestic pigs and native boar in the evacuation zone after the accident and was detected in 15 samples across a broad geographic area. Our MIG-seq analysis revealed genetic structure of boar was significantly different between boar inhabiting the eastern (including FEZ. i.e., East clade) and western (i.e., West clade) regions in Fukushima prefecture. In addition, we investigated the relationships between boar dispersal and Cesium (Cs)-137 activity concentrations in boar muscle using MIG-seq genetic data in Nihonmatsu city, located in the central-northern region of Fukushima. High Cs-137 activity concentrations, exceeding 1000 Bq/kg, in boar muscle had a significantly high probability of belonging to the East clade within localized regions. Thus, our results provide evidence of the spatial scale of dispersal of individuals or offspring of boar from the FEZ. Results of this research also indicate that dispersal of individuals between areas with different Cs-137 contamination levels is one of the biggest factors contributing to variation in Cs-137 activity concentration in boar muscle within localized regions.
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  • 文章类型: Journal Article
    需要更好地理解生物材料中的DNA降解。洗过的衣服上的血迹会受到洗涤程序的干扰,有时转移到其他织物上,通常有潜在的血迹,通常有显著降解的DNA。样品(有血迹的棉织物)分为六个主要组,根据水温(95、60和30°C)的洗涤方法和洗涤剂的使用。完成洗涤过程后,样本储存一段时间(1天至6个月),随后进行分析.使用标准方案和商业试剂盒进行分析以测量经处理的样品中的剩余DNA量(浓度)和DNA降解指数。我们的结果表明,高洗涤温度(60和95°C)和洗涤剂的应用对DNA降解具有协同作用,而在30°C时,这种效果是不存在的。此外,在洗涤后约一个月观察到洗涤剂对加速DNA降解的影响。洗涤剂的这种延迟效应在目前的文献数据中没有解释。为了从血迹中获得最佳结果,我们建议从犯罪事件和肇事者试图清洁到实验室分析的时间应少于1个月。
    DNA degradation in biological material needs to be better understood. Bloodstains on washed clothing are disturbed by washing procedures, sometimes transferred to other fabrics, often with latent bloodstains and usually with significantly degraded DNA. The samples (cotton fabric with bloodstains) are divided into six main groups, depending on the washing method regarding water temperature (95, 60, and 30 °C) and the detergent use. After completing the washing process, samples were stored for a certain period (1 day to 6 months) and subsequently analyzed. Analyses were performed using standard protocols and commercial kits to measure the remaining DNA quantity (concentration) and DNA degradation index in the processed samples. Our results revealed that the high washing temperature (60 and 95 °C) and the application of detergent have a synergic action on DNA degradation, while at 30 °C this effect is absent. Furthermore, the effect of detergent on accelerated DNA degradation is observed about a month after the washing. This delayed effect of detergent has no explanation in current literature data. To obtain optimal results from the bloodstains, we recommended that the period from the crime event and attempted cleaning by a perpetrator to the laboratory analysis should be less than 1 month.
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  • 文章类型: Case Reports
    作者介绍了一个部分葡萄胎的案例,其中DNA分析(STR-短串联重复基因分型)显示了一个具有XXXY性腺补体的三性单性四倍体基因组组成。这一遗传学发现在临床病理上与部分葡萄胎相关,尽管与典型的相比很少见,二重单性三倍体部分痣。遗传分析明确证实了部分葡萄胎的可疑诊断。为了排除选择性终止妊娠后,磨牙妊娠代表保留的受孕产物的可能性,比较了磨牙妊娠和先前受孕产物的STR谱。短串联重复序列基因分型是部分葡萄胎鉴别诊断的一种有用的分子遗传学方法,临床病理结果经常模棱两可。
    The authors present a case of a partial hydatidiform mole where DNA analysis (STR - short tandem repeat genotyping) showed a triandric monogynic tetraploid genome composition with a XXXY gonosomal complement. This genetic finding clinicopathologically correlates with a partial hydatidiform mole, although it is rare in comparison with the typical, diandric monogynic triploid partial moles. The genetic analysis definitively confirmed the suspected diagnosis of a partial mole. To exclude the possibility that molar pregnancy represented retained products of conception after elective pregnancy termination, STR profiles from molar pregnancy and previous products of conception were compared. Short tandem repeats genotyping is a useful molecular genetic method in the differential diagnosis of partial hydatidiform moles, where clinical-pathological findings are frequently ambiguous.
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  • 文章类型: Journal Article
    分析犯罪现场的所有生物证据是严肃的时刻,预算,和劳动力约束。因此,选择有效的证据对于有效筛查至关重要。ABO血型是一种标记,可以作为在刑事案件中识别调查线索的有效证据。使用血清学方法常规鉴定ABO血型仅用于血液,难以应用于其他体液。通过分析代表每个血型的单核苷酸多态性(SNP)进行ABO基因分型。然而,这种方法很耗时,贵,需要精密的仪器.在这项研究中,我们使用环介导等温扩增(LAMP)和多重实时聚合酶链反应(PCR)开发了快速ABO基因分型方法。选择ABO基因中的三个SNP位点(nt261、526和803)来对ABO基因型进行分类。对于特异性测试,我们对60份唾液样本进行了测序,以确认基因分型。我们进行了实验,使用两种扩增方法应用ABO基因分型,使用棉签和滤纸模拟法医样品。因此,使用LAMP,在恒温(65℃)30min内成功鉴定出6种ABO基因型。此外,通过使用多重实时PCR,不仅可以检测主要组,还可以检测ABO基因型的亚组(例如顺式AB)。使用新方法的扩增结果与测序结果一致。因此,这些ABO基因分型方法有望在犯罪现场成功有效地选择有效证据。
    Analyzing all biological evidence at a crime scene presents serious time, budget, and labor constraints. Therefore, selecting valid evidence is crucial for efficient screening. The ABO blood group is a marker that can serve as valid evidence for identifying investigative leads in criminal case. Conventional identification of ABO blood groups using serological methods has only been for blood and is difficult to apply to other body fluids. ABO genotyping was conducted by analyzing single nucleotide polymorphisms (SNP) representative of each blood group. However, this method is time-consuming, expensive, and requires sophisticated instruments. In this study, we developed rapid ABO genotyping method using loop-mediated isothermal amplification (LAMP) and multiplex real-time polymerase chain reaction (PCR). Three SNP sites in the ABO gene (nt 261, 526, and 803) were selected to classify the ABO genotypes. For the specificity test, we performed sequencing of 60 saliva samples to confirm that the genotyping. We conducted experiments to apply ABO genotyping using two amplification methods to mock forensic sample using cotton swab and filter paper. As a result, using LAMP, we successfully identified six ABO genotypes within 30 min at a constant temperature (65 ℃). Moreover, by using multiple real-time PCR, it was possible to detect not only the major group but also the subgroup of the ABO genotype (ex. cis-AB). The amplification results using the new methods were in concordance with the sequencing results. Therefore, these ABO genotyping methods are expected to select valid evidence successfully and efficiently at the crime scene.
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