Polymerase chain reaction

聚合酶链反应
  • 文章类型: Case Reports
    单纯疱疹性脑炎(HSVE)是一种潜在致命的传染性中枢神经系统(CNS)疾病。因此,早期发现是决定案件命运的关键。临床病史和检查,脑计算机断层扫描,动态对比增强磁共振成像(DCE-MRI),腰椎穿刺已经被用来建立诊断。本报告描述了一例HSVE,伴有低细胞脑脊液(CSF)和罕见的记忆障碍。然而,MRI结果与HSVE一致,和CSFPCR检测对治疗有反应的HSV-1DNA呈阳性。我们通常建议患者尽快开始抗病毒治疗,以避免并发症。
    Herpes simplex encephalitis (HSVE) is a potentially fatal infectious central nervous system (CNS) disorder. Thus, early detection is critical in determining the case\'s fate. Clinical history and examination, brain computed tomography, dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI), and lumbar puncture have been used to establish a diagnosis. This report describes a case of HSVE with hypocellular cerebrospinal fluid (CSF) and an uncommon form of memory impairment. However, MRI results were consistent with HSVE, and CSF PCR tested positive for HSV-1 DNA that responded to treatment. We routinely advise patients to begin antiviral therapy as soon as possible to avoid complications.
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  • 文章类型: Systematic Review
    背景:猫相关的嗜血支原体(血液原虫)被认为通过两种主要机制传播:(1)通过战斗直接传播和(2)由猫蚤(Ctenocephalidesfelis)传播。虽然C.felis的传输效率似乎很低,大多数手稿都集中在野生跳蚤中的血原虫的患病率,并报告了非常低(<3%)或高(>26%)的患病率。因此,我们旨在评估样品处理和PCR方法对C.felis血浆感染患病率的影响。
    方法:对PubMed文章的系统审查确定了13份手稿(1,531只跳蚤/跳蚤池)符合纳入标准(对从猫收集的C.felis上的>1份血浆进行PCR)。使用ROBINS-E工具评估偏倚风险。在这些手稿的R中进行的荟萃分析发现,不洗涤样品和一组常见的16SrRNA引物首次发表在Jensen等人。2001年与血血浆患病率增加有关。为了评估洗涤对新收集的跳蚤的影响,我们评估了20个5个C.felis池的血浆状态,其中一半被洗了,一半没有洗。
    结果:跳蚤冲洗并不影响血血浆的检测,而是扩增了螺血浆。用Jensen等人评估非特异性扩增。2001引物,对67例C.felis样品(34%先前报道的血血浆感染)进行PCR和测序。通过这种方法,仅在3%的样本中检测到血浆.在剩下的“血支原体感染”跳蚤中,PCR扩增螺旋体或其他细菌。
    结论:因此,我们得出的结论是,在C.felis中的血浆感染是罕见的,未来的跳蚤流行研究应该对所有阳性扩增子进行测序以验证PCR特异性。有必要进一步研究猫相关的血血浆传递的替代方法以及C.felis维持血血浆感染的能力。
    BACKGROUND: Feline-associated hemotropic Mycoplasma (hemoplasmas) are believed to be transmitted by two primary mechanisms: (1) direct transmission via fighting and (2) vector-borne transmission by the cat flea (Ctenocephalides felis). While the efficiency of transmission by C. felis appears low, most manuscripts focus on the prevalence of hemoplasmas in wild-caught fleas and report either a very low (< 3%) or a high (> 26%) prevalence. Therefore, we aimed to assess the influence of sample processing and PCR methods on C. felis hemoplasma infection prevalence.
    METHODS: A systemic review of PubMed articles identified 13 manuscripts (1,531 fleas/flea pools) that met the inclusion criteria (performed PCR for >1 hemoplasma on C. felis collected from cats). Risk of bias was assessed utilizing the ROBINS-E tool. Meta-analysis performed in R of these manuscripts found that not washing samples and a common set of 16S rRNA primers first published in Jensen et al. 2001 were associated with increased hemoplasma prevalence. To evaluate the influence of washing on newly collected fleas, we assessed the hemoplasma status of 20 pools of 5 C. felis each, half of which were washed and half not washed.
    RESULTS: Flea washing did not influence the detection of hemoplasma but instead amplified Spiroplasma. To assess non-specific amplification with the Jensen et al. 2001 primers, 67 C. felis samples (34% previously reported hemoplasma infected) were subject to PCR and sequencing. By this method, hemoplasma was detected in only 3% of samples. In the remaining \"hemoplasma infected\" fleas, PCR amplified Spiroplasma or other bacteria.
    CONCLUSIONS: Therefore, we concluded that hemoplasma infection in C. felis is rare, and future flea prevalence studies should sequence all positive amplicons to validate PCR specificity. Further investigation of alternative methods of feline-associated hemoplasma transmission and the ability of C. felis to maintain hemoplasma infection is necessary.
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  • 文章类型: Journal Article
    背景:脑结核瘤(BT)的诊断有时具有挑战性。在这里,我们提出了一个案例系列来评估联合诊断方法,包括抗酸杆菌(AFB)染色,聚合酶链反应(PCR),基因Xpert,和组织病理学,结核瘤组织标本(TTSs)。患者和方法:共有16例患者(11例人类免疫缺陷病毒[HIV]阳性,本研究包括通过TTS的联合诊断方法证实的5个HIV阴性)和BT。临床数据,包括临床症状,实验室测试,神经影像学特征,组织病理学,治疗,和预后,对所有患者进行评估。结果:男性10例,女性6例(范围,18-73岁)。11例和10例患者的抗酸杆菌染色和TTSsPCR均为阳性,分别。TTS基因Xpert的敏感性为(80.0%;8/10)。9例(56.3%;9/16)患者通过组织病理学诊断为BT。在接受抗结核治疗后,12例(75.0%;12/16)患者在临床上有相当大的改善。结论:TTS的联合诊断方法可提高BT的诊断效率。
    Background: The diagnosis of brain tuberculoma (BT) is sometimes challenging. Herein, we presented a case series to evaluate the combined-diagnostic methods, including acid-fast bacilli (AFB) stain, polymerase chain reaction (PCR), Gene Xpert, and histopathology, of tuberculoma tissue specimens (TTSs). Patients and Methods: A total of 16 patients (11 human immunodeficiency virus [HIV]-positive, 5 HIV-negative) with BT confirmed by combined-diagnostic methods of TTS were included in this study. Clinical data, including clinical symptoms, laboratory tests, neuroimaging features, histopathology, treatment, and prognosis, were assessed in all patients. Results: There were 10 male and 6 female patients (range, 18-73 years). Acid-fast bacilli stain and PCR of TTSs were positive in 11 and 10 patients, respectively. The sensitivity of Gene Xpert of TTSs was (80.0%; 8/10). Nine (56.3%; 9/16) patients were diagnosed with BT by histopathology. After receiving antituberculosis treatment, 12 (75.0%; 12/16) patients improved clinically to a considerable extent. Conclusions: The combined-diagnostic methods of TTS may improve the diagnostic efficiency of BT.
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  • 文章类型: Case Reports
    曲霉病是中枢神经系统中罕见的感染,死亡率高于95%。早期诊断具有挑战性且至关重要。在这份报告中,我们介绍了一名六岁女性,患有剧烈头痛并伴有左偏瘫的病例,凝视偏差,水平眼球震颤,呕吐粘液内容物五次。经过几次接近,脑脊液PCR结果为曲霉属阳性。,然后以2.6mg/kg/天的两性霉素B开始管理,并管理使用伏立康唑。她活了下来,在她第一次入院两年后,她患有脑曲霉病后遗症。改进的一个领域是机构外研究的请求和交付之间的协调。在这种情况下,患者的母亲没有按时报告分析结果,延迟诊断。
    Aspergillosis is an infrequent infection in the Central Nervous System with a mortality rate higher than 95 %. Early diagnosis is challenging and crucial. In this report, we present the case of a six-year-old female with an intense headache accompanied by left hemiparesis, gaze deviation, horizontal nystagmus, and vomiting of mucous content on five occasions. After several approaches, a cerebrospinal fluid PCR resulted positive for Aspergillus spp., and then management started with amphotericin B at 2.6 mg/kg/day and was managed to have voriconazole. She survived, and two years after her first hospital admission, she suffered from cerebral aspergillosis sequelae. An area of improvement is the coordination between the request and delivery of studies outside the institution. In this case, the patient´s mother did not report the analysis results on time, delaying the diagnosis.
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  • 文章类型: Letter
    暂无摘要。
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  • 文章类型: Case Reports
    单侧急性特发性黄斑病变(UAIM)是一种罕见的疾病,在流感样疾病后可能导致年轻人的单侧视力丧失。偶尔,它与手足口病(HFMD)有关,通常未被诊断。在这里,我们报告了一例UAIM与HFMD合并急性肠道病毒感染相关的病例。根据使用多模态诊断成像的临床发现,包括眼底图像,光学相干层析成像,荧光血管造影术,以及泛肠道病毒RNA聚合酶链反应的血清学测试,我们详细介绍了UAIM合并HFMD相关脉络膜炎的临床过程,并推测其发病机制。这项研究可以提醒每位医生UAIM在手足口病中可能导致的视力丧失,将患者转诊到眼科检查对消除潜在的视力损害很重要。
    Unilateral acute idiopathic maculopathy (UAIM) is a rare disease that may cause unilateral vision loss in young adults after a flu-like illness. Occasionally, it is associated with hand-foot-mouth disease (HFMD) and is often underdiagnosed. Herein, we report a case with characteristics of UAIM associated with HFMD with acute enteroviral infection. On the basis of the clinical findings using multimodality diagnostic imaging, including fundus image, optical coherence tomography, fluorescence angiography, and serological test for pan-enterovirus RNA polymerase chain reaction, we detailed the clinical course and postulated the pathogenesis of UAIM with choroiditis associated with HFMD. This study could remind every doctor of the potential visual loss caused by UAIM in HFMD, and referring patients to the ophthalmologic survey is important to eliminate potential visual impairment.
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  • 文章类型: Journal Article
    背景:2020年,国家和领土流行病学家委员会(CSTE)百日咳病例定义进行了修改;主要变化是将PCR阳性病例分类为已确认,无论咳嗽持续时间。通过七个地点的增强百日咳监测(EPS)和国家法定疾病监测系统(NNDSS)报告的百日咳数据用于评估新病例定义的影响。
    方法:我们比较了2020年咳嗽发作的EPS病例数与根据先前(2014年)CSTE病例定义报告的病例数。为了在全国范围内评估变化的影响,根据2020年CSTE病例定义新报告的EPS病例比例应用于2020年NNDSS数据,以估计全国新增病例数.
    结果:根据2020年病例定义,在2020年报告给EPS的442例确诊和可能病例中,有42例(9.5%)是新报告病例。将这一比例应用于2020年全国报告的6124例确诊和可能病例,我们估计新定义增加了582例。如果案例定义没有改变,2020年报告的病例将比2019年减少70%;观察到的减少是67%。
    结论:尽管在COVID-19病例中报告的百日咳病例大幅减少,但我们的数据显示,与以前的病例定义相比,2020年百日咳病例定义的改变导致了额外的病例报告,为公共卫生干预措施提供更多机会,例如预防密切接触者。
    BACKGROUND: In 2020, the Council of State and Territorial Epidemiologists (CSTE) pertussis case definition was modified; the main change was classifying polymerase chain reaction (PCR)-positive cases as confirmed, regardless of cough duration. Pertussis data reported through Enhanced Pertussis Surveillance (EPS) in 7 sites and the National Notifiable Diseases Surveillance System (NNDSS) were used to evaluate the impact of the new case definition.
    METHODS: We compared the number of EPS cases with cough onset in 2020 to the number that would have been reported based on the prior (2014) CSTE case definition. To assess the impact of the change nationally, the proportion of EPS cases newly reportable under the 2020 CSTE case definition was applied to 2020 NNDSS data to estimate how many additional cases were captured nationally.
    RESULTS: Among 442 confirmed and probable cases reported to EPS states in 2020, 42 (9.5%) were newly reportable according to the 2020 case definition. Applying this proportion to the 6124 confirmed and probable cases reported nationally in 2020, we estimated that the new definition added 582 cases. Had the case definition not changed, reported cases in 2020 would have decreased by 70% from 2019; the observed decrease was 67%.
    CONCLUSIONS: Despite a substantial decrease in reported pertussis cases in the setting of coronavirus disease 2019 (COVID-19), our data show that the 2020 pertussis case definition change resulted in additional case reporting compared with the previous case definition, providing greater opportunities for public health interventions such as prophylaxis of close contacts.
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  • 文章类型: Journal Article
    弓形虫是一种普遍存在的寄生原虫,可能是神经和精神疾病的重要原因。这项基于病例对照登记的研究的目的是评估在马赞达兰省伊朗国家弓形虫病登记中心通过吸毒企图自杀的受试者和对照组中弓形虫感染的患病率和相关危险因素,伊朗北部。基线数据是使用问卷从参与者那里收集的,并从每个人身上采集血液样本。血浆准备用于血清学分析,而血沉棕黄层用于分子分析。在282个人中(147例自杀未遂[SA]和135例对照),42.9%的患者和16.3%的对照受试者抗弓形虫免疫球蛋白G(IgG)阳性,但所有参与者的弓形虫DNA和抗弓形虫免疫球蛋白M均为阴性。20-30岁年龄组中SA的IgG血清阳性率是对照组的3.22倍(p<0.001)。这些发现表明,SA中潜伏性弓形虫感染明显高于健康个体,至少在研究区域,表明潜在弓形虫病与SA之间存在潜在关联。需要进一步的研究来阐明弓形虫与世界不同人群和地区之间自杀之间的潜在联系。
    Toxoplasma gondii is a ubiquitous parasitic protozoan that may be an important cause of neurological and psychiatric diseases. The purpose of this case-control registry-based study was to evaluate the prevalence of T. gondii infection and related risk factors among subjects who attempted suicide by drug use and a control group at the Iranian National Registry Center for Toxoplasmosis in Mazandaran Province, northern Iran. Baseline data were collected from participants using a questionnaire, and a blood sample was taken from each individual. The plasma was prepared for serological analysis, whereas the buffy coat was used for molecular analysis. Out of 282 individuals (147 cases with suicide attempters [SA] and 135 controls), 42.9% of patients and 16.3% of control subjects were positive for anti-Toxoplasma immunoglobin G (IgG), but all participants were negative for T. gondii DNA and anti-Toxoplasma immunoglobin M. Based on multiple logistic regressions, IgG seropositivity in SA in the age group of 20-30 years was 3.22 times higher than that in the control group (p < 0.001). These findings suggest that latent T. gondii infection among SA is significantly higher than that in healthy individuals, indicating a potential association between latent toxoplasmosis and SA at least in the studied area. Further research is needed to shed light on the potential association between T. gondii and suicide among different populations and areas of the world.
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  • 文章类型: Case Reports
    一个成年马的临床病例,溃疡性,增殖性,提出了肩部区域皮肤(肿瘤)的脓性肉芽肿病。肿块呈肉芽肿状和火山口状,有血清血排出物和有干酪样物质的瘘管。手术切除肿瘤,送实验室诊断。组织病理学使用Grocott-Gomori次甲基胺银染进行。坏死物质的存在,纤维化,浸润细胞,和棕色的菌丝,Pythium属成员的特征,被观察到。为了识别感染物种,进行了用于扩增ITS-1的常规PCR。组织病理学和PCR测试证实了与美国和中美洲以前的记录密切相关的煤腐菌菌株感染。我们的报告代表了墨西哥第一个分子确认的马化脓症病例。
    A clinical case of an adult horse with invasive, ulcerative, proliferative, pyogranulomatous disease of the skin (tumor) in the shoulder region is presented. The mass had a granulomatous and crater-shaped appearance, with serosanguinous discharge and the presence of fistulas with caseous material. The tumor was removed by surgery and sent to the laboratory for diagnosis. Histopathology was performed using Grocott-Gomori methenamine silver stain. The presence of necrotic material, fibrosis, infiltrated cells, and brown-colored hyphae, characteristic of members of the genus Pythium, were observed. To identify the infecting species, conventional PCRs for the amplification of the ITS-1 was carried out. Histopathological and PCR tests confirmed infection by a Pythium insidiosum strain closely associated with previous records from the US and Central America. Our report represents the first molecularly confirmed case of equine pythiosis in Mexico.
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  • 文章类型: Journal Article
    背景:河马果蝇是吸血节肢动物,可以传播病原微生物,因此是病原体如巴尔通体的潜在载体。这些革兰氏阴性菌可在人类和动物中引起轻度至重度的临床体征;因此,监测巴尔通菌属。虱虫种群的患病率似乎是人畜共患风险评估的有用前提。
    方法:使用方便的抽样,我们从四个ked物种(Lipoptenacervi,n=22;fortisetosalipoptenafortisetosa,n=61;龙舌兰,n=12;马匹,n=8)和萨克森州联邦州的M.ovinus(n=10)的p,德国。通过聚合酶链反应(PCR)筛选所有样品中的巴尔通菌。DNA,靶向柠檬酸合酶基因(gltA)。随后,靶向五个以上基因的PCRs(16S,ftsZ,nuoG,对揭示的gltA基因型的代表进行了ribC和rpoB),并对所有PCR产物进行测序,以准确鉴定巴尔通体(亚)种。
    结论:巴尔通体的总体检出率。是100.0%,59.1%,M.ovinus中的24.6%和75.0%,L.cervi,L.fortisetosa和马匹,分别。所有已鉴定的Bartonellae都属于Bartonellaschanenbuchensis复合体。我们的数据支持对该组(亚)种状态的重新分类,因此,我们得出结论,B.schoenbuchensis的几个基因型被检测到,包括巴尔通菌亚种。墨罗哈木和巴尔通菌亚种。schoenbuchensis,两者先前都验证了人畜共患的潜力。广泛的PCR分析显示,必须采用多种PCR方法来正确鉴定反刍动物相关的bartonellae。
    Hippoboscid flies are bloodsucking arthropods that can transmit pathogenic microorganisms and are therefore potential vectors for pathogens such as Bartonella spp. These Gram-negative bacteria can cause mild-to-severe clinical signs in humans and animals; therefore, monitoring Bartonella spp. prevalence in louse fly populations appears to be a useful prerequisite for zoonotic risk assessment.
    Using convenience sampling, we collected 103 adult louse flies from four ked species (Lipoptena cervi, n = 22; Lipoptena fortisetosa, n = 61; Melophagus ovinus, n = 12; Hippobosca equina, n = 8) and the pupae of M. ovinus (n = 10) in the federal state of Saxony, Germany. All the samples were screened by polymerase chain reaction (PCR) for Bartonella spp. DNA, targeting the citrate synthase gene (gltA). Subsequently, PCRs targeting five more genes (16S, ftsZ, nuoG, ribC and rpoB) were performed for representatives of revealed gltA genotypes, and all the PCR products were sequenced to identify the Bartonella (sub)species accurately.
    The overall detection rates for Bartonella spp. were 100.0%, 59.1%, 24.6% and 75.0% in M. ovinus, L. cervi, L. fortisetosa and H. equina, respectively. All the identified bartonellae belong to the Bartonella schoenbuchensis complex. Our data support the proposed reclassification of the (sub)species status of this group, and thus we conclude that several genotypes of B. schoenbuchensis were detected, including Bartonella schoenbuchensis subsp. melophagi and Bartonella schoenbuchensis subsp. schoenbuchensis, both of which have previously validated zoonotic potential. The extensive PCR analysis revealed the necessity of multiple PCR approach for proper identification of the ruminant-associated bartonellae.
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