Specificity

特异性
  • 文章类型: Journal Article
    伯氏柯西拉(C.burnetii)感染的牲畜和野生动植物在流行病学上与人类Q热暴发有关。尽管人畜共患的威胁越来越大,对野生动物中的考氏杆菌病的知识仍然有限,需要进行研究以了解其流行病学作用。在C.Burnetii流行地区,据报道,壁虱可以携带和传播C.burnetii,并可能作为野生动物栖息地感染风险的指标。因此,这项研究的目的是比较检测蜱中的C.burnetiiDNA的分子技术。
    总共,使用常规PCR(cPCR)和两种现场友好技术:Biomeme的C.burnetiiqPCRGo-strip(Biomeme)和新的C.burnetiiPCR高分辨率熔解(PCR-HRM)分析测定。结果进行了评估,在没有黄金标准测试的情况下,使用贝叶斯潜在类别分析(BLCA)来表征C.burnetii阳性蜱的比例,并估计三种测试的敏感性(Se)和特异性(Sp)。
    最终的BLCA模型包括主要效应,并估计PCR-HRM具有最高的Se(86%;95%可信区间:56-99%),其次是生物eme(Se=57%;95%可信区间:34-90%),cPCR的估计硒最低(24%,95%可信区间:10-47%)。所有三种测定的特异性估计范围为94至98%。基于模型,估计有16%的蜱有C.burnetiiDNA存在。
    这些结果反映了肯尼亚北部B.Burnetii的地方性,并显示了PCR-HRM测定用于壁虱B.Burnetii监测的前景。使用蜱和野生动物样本的进一步研究将增强对蜱在Q热中的流行病学作用的理解。
    UNASSIGNED: Coxiella burnetii (C. burnetii)-infected livestock and wildlife have been epidemiologically linked to human Q fever outbreaks. Despite this growing zoonotic threat, knowledge of coxiellosis in wild animals remains limited, and studies to understand their epidemiologic role are needed. In C. burnetii-endemic areas, ticks have been reported to harbor and spread C. burnetii and may serve as indicators of risk of infection in wild animal habitats. Therefore, the aim of this study was to compare molecular techniques for detecting C. burnetii DNA in ticks.
    UNASSIGNED: In total, 169 ticks from wild animals and cattle in wildlife conservancies in northern Kenya were screened for C. burnetii DNA using a conventional PCR (cPCR) and two field-friendly techniques: Biomeme\'s C. burnetii qPCR Go-strips (Biomeme) and a new C. burnetii PCR high-resolution melt (PCR-HRM) analysis assay. Results were evaluated, in the absence of a gold standard test, using Bayesian latent class analysis (BLCA) to characterize the proportion of C. burnetii positive ticks and estimate sensitivity (Se) and specificity (Sp) of the three tests.
    UNASSIGNED: The final BLCA model included main effects and estimated that PCR-HRM had the highest Se (86%; 95% credible interval: 56-99%), followed by the Biomeme (Se = 57%; 95% credible interval: 34-90%), with the estimated Se of the cPCR being the lowest (24%, 95% credible interval: 10-47%). Specificity estimates for all three assays ranged from 94 to 98%. Based on the model, an estimated 16% of ticks had C. burnetii DNA present.
    UNASSIGNED: These results reflect the endemicity of C. burnetii in northern Kenya and show the promise of the PCR-HRM assay for C. burnetii surveillance in ticks. Further studies using ticks and wild animal samples will enhance understanding of the epidemiological role of ticks in Q fever.
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  • 文章类型: Journal Article
    目标:为了解决由于筛查工具有限而导致的痴呆及时诊断的差距,我们调查了Hellocog的有效性和可靠性,基于片剂的计算机化神经心理学测试用于筛查痴呆症。Hellocog上的概率分数越高,患痴呆症的可能性越高。
    方法:本研究包括100名痴呆患者和100名认知正常的人,他们年龄在60岁或以上,没有其他主要的精神病患者,神经学,或医疗条件。他们在神经心理学家的监督下,在平板电脑上使用Hellocog。为了确定测试-重测可靠性,20在4周后再次服用了Hellocog。使用接受者操作员特征(ROC)分析评估诊断性能。
    结果:Hellocog显示出足够的内部一致性(Cronbach'salpha=0.69)和良好的重测可靠性(组内相关系数=0.86,p<0.001)。患有痴呆症的参与者在Hellocog上的得分高于认知正常的参与者(p<0.001),证实了其高标准的有效性。与小型精神状态检查(MMSE)评分和建立阿尔茨海默病神经心理学评估数据库(CERAD-TS)注册联盟的总分密切相关,突出了Hellocog的并发有效性。Hellocog痴呆的ROC曲线下面积非常好(0.971),与MMSE和CERAD-TS相当。对痴呆的敏感性和特异性分别为0.945%和0.872%,分别,略优于MMSE和CERAD-TS。
    结论:Hellocog作为自我管理的痴呆筛查的有效和可靠的工具,有望改善痴呆症的早期发现。
    OBJECTIVE: To address the gap in timely diagnosis of dementia due to limited screening tools, we investigated the validity and reliability of the Hellocog, computerized neuropsychological test based on tablets for screening dementia. The higher the probability score on the Hellocog, the higher the likelihood of dementia.
    METHODS: This study included 100 patients with dementia and 100 individuals with normal cognition who were aged 60 years or older and free of other major psychiatric, neurological, or medical conditions. They administered the Hellocog on a tablet under the supervision of a neuropsychologist. To determine test-retest reliability, 20 took the Hellocog again after 4 weeks. Diagnostic performance was assessed using the receiver operator characteristics (ROC) analysis.
    RESULTS: The Hellocog showed adequate internal consistency (Cronbach\'s alpha=0.69) and good test-retest reliability (intraclass correlation coefficient=0.86, p<0.001). Participants with dementia scored higher on the Hellocog than those with normal cognition (p<0.001), confirming its high criterion validity. Strong correlations with the Mini-Mental Status Examination (MMSE) score and the total score of the Consortium to Establish a Registry for Alzheimer\'s Disease Neuropsychological Assessment Battery (CERAD-TS) highlight the concurrent validity of the Hellocog. The area under the ROC curve for dementia of the Hellocog was excellent (0.971) and comparable to that of the MMSE and CERAD-TS. The sensitivity and specificity for dementia were 0.945 and 0.872%, respectively, which were slightly better than those of the MMSE and CERAD-TS.
    CONCLUSIONS: Hellocog stands out as a valid and reliable tool for self-administered dementia screening, with promise for improving early detection of dementia.
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  • 文章类型: Journal Article
    ELISA测定通常用于赛车实验室的药物筛选,但已知特异性有限。不准确的ELISA筛选结果通常是通过非特异性抗体相互作用或由于样品降解导致的显色材料在样品孔中的保留而产生的。虽然药物阳性的确认可以通过质谱来实现,ELISA筛查结果不准确的随访意味着员工时间和试剂的不必要成本.在使用夹心ELISA测定法进行rhEPO筛选的情况下尤其如此,其中确认方法最多需要3天才能执行。虽然大多数赛车实验室购买商业ELISA试剂盒,这些产品可以定制,以提供增加的特异性,以增强阳性样品的筛选。商业夹心ELISA试剂盒的特异性可以通过多种机制来提高,包括添加竞争性分析物特异性抗体,替代捕获抗体或通过在有和没有捕获抗体的情况下进行ELISA分析。困难基质如犬尿中的非特异性信号也可以通过在添加样品之前将BSA溶液添加到ELISA板中而减少。
    ELISA assays are commonly used for drug screening by racing laboratories but are known to suffer from limited specificity. Inaccurate ELISA screening results are typically produced by non-specific antibody interactions or by the retention of chromogenic material in the sample well due to sample degradation. While confirmation of drug positives can be achieved by mass spectrometry, the follow-up of inaccurate ELISA screening results represents an unnecessary cost in staff time and reagents. This is particularly true in the case of rhEPO screening using sandwich ELISA assays, where the confirmation method requires up to 3 days to perform. While most racing laboratories purchase commercial ELISA kits, these products can be customised to provide increased specificity for enhanced screening of positive samples. The specificity of commercial sandwich ELISA kits can be improved by a variety of mechanisms including the addition of competing analyte specific antibodies, substitution of capture antibodies or by performing ELISA analysis with and without capture antibodies. Non-specific signals in difficult matrices such as canine urine can also be reduced by the addition of BSA solutions to the ELISA plate prior to the addition of samples.
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  • 文章类型: Journal Article
    目的:心理健康量表(MHI-5)经常用作情绪和焦虑症的筛查工具。然而,很少有基于人群的研究针对按照当前诊断标准评估疾病的诊断工具进行验证.
    方法:在第三次荷兰心理健康调查和发病率研究(NEMESIS-3)中,一项代表性的基于人群的成人研究(N=6194;年龄:18-75岁),MHI-5在过去的一个月中用于测量一般的精神疾病.情绪的存在(重度抑郁症,持续性抑郁障碍,或躁郁症)和焦虑症(恐慌症,广场恐惧症,社交恐惧症,根据《精神障碍诊断和统计手册-5》,对过去一个月的综合国际诊断访谈3.0进行了稍微修改的评估。
    结果:MHI-5在区分有和没有情绪障碍的人方面非常出色,焦虑症,和任何情绪或焦虑症。与情绪障碍的最高敏感性和最高特异性相关的临界值≤68,对于焦虑症或任何情绪或焦虑症,临界值≤76。
    结论:当诊断性访谈太耗时时,MHI-5可以识别普通人群中当前情绪或焦虑症的高风险个体。
    OBJECTIVE: The Mental Health Inventory (MHI-5) is frequently used as a screener for mood and anxiety disorders. However, few population-based studies have validated it against a diagnostic instrument assessing disorders following current diagnostic criteria.
    METHODS: Within the third Netherlands Mental Health Survey and Incidence Study (NEMESIS-3), a representative population-based study of adults (N = 6194; age: 18-75 years), the MHI-5 was used to measure general mental ill-health in the past month. Presence of mood (major depressive disorder, persistent depressive disorder, or bipolar disorder) and anxiety disorders (panic disorder, agoraphobia, social phobia, or generalized anxiety disorder) in the past month was assessed with a slightly modified version of the Composite International Diagnostic Interview 3.0 per the Diagnostic and Statistical Manual of Mental disorders-5.
    RESULTS: The MHI-5 was good to excellent at distinguishing people with and without a mood disorder, an anxiety disorder, and any mood or anxiety disorder. The cut-off value associated with the highest sensitivity and highest specificity for mood disorder was ≤68, and ≤76 for an anxiety disorder or any mood or anxiety disorder.
    CONCLUSIONS: The MHI-5 can identify individuals at high risk of a current mood or anxiety disorder in the general population when diagnostic interviews are too time consuming.
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  • 文章类型: Journal Article
    镰刀菌冠腐病(FCR),由镰刀菌引起。,是小麦种植区的毁灭性疾病。以前的研究表明,FCR是由同时感染禾谷镰刀菌引起的,F.假赤霉,湖北省F.proliferatum和F.writicillioides,中国。在这项研究中,开发了一种同时检测F.graminearumDNA的方法,F.假赤霉,可以有效区分它们的F.proliferatum和F.roticillioides。这四个镰刀菌的全基因组序列比较。进行,并设计了20bp序列作为通用上游引物。还设计了每种病原体的特异性下游引物,每个病原体产生206、482、680和963bp的扩增子,分别。多重PCR专门鉴定了禾谷镰刀菌,F.假赤霉,F.增殖和轮虫,但不来自其他46种病原体,目标病原体的检测限约为100pg/μl。此外,我们使用优化的多重PCR方法准确地确定了小麦样品中的FCR病原体种类。这些结果表明,本研究建立的多重PCR方法可以高效、快速地鉴定禾谷菌株。F.假赤霉,F.增殖,和F.verticillioides,为及时、有针对性地预防和控制FCR提供技术支持。
    Fusarium crown rot (FCR), caused by Fusarium spp., is a devastating disease in wheat growing areas. Previous studies have shown that FCR is caused by co-infection of F. graminearum, F. pseudograminearum, F. proliferatum and F. verticillioides in Hubei Province, China. In this study, a method was developed to simultaneously detected DNAs of F. graminearum, F. pseudograminearum, F. proliferatum and F. verticillioides that can efficiently differentiate them. Whole genome sequence comparison of these four Fusarium spp. was performed and a 20 bp sequence was designed as an universal upstream primer. Specific downstream primers of each pathogen was also designed, which resulted in a 206, 482, 680, and 963 bp amplicon for each pathogen, respectively. Multiplex PCR specifically identified F. graminearum, F. pseudograminearum, F. proliferatum and F. verticillioides but not from other 46 pathogens, and the detection limit of target pathogens is about 100 pg/μl. Moreover, we accurately determined the FCR pathogen species in wheat samples using the optimized multiplex PCR method. These results demonstrate that the multiplex PCR method established in this study can efficiently and rapidly identify F. graminearum, F. pseudograminearum, F. proliferatum, and F. verticillioides, which should provide technical support for timely and targeted prevention and control of FCR.
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  • 文章类型: Journal Article
    背景:指甲积聚酒精代谢产物,乙基葡糖苷酸(ETG),和大麻代谢物,羧基-δ-9-THC超过3-6个月。很少有研究检查指甲毒理学测试的敏感性和特异性,以及指甲测试与自我报告的酒精和大麻使用之间的协议。
    方法:在一项正在进行的临床试验中,1101名退伍军人完成了最初的电话问卷调查,然后被要求邮寄指甲剪报以进行物质使用分析。与酒精使用模式(AUDIT-C)和物质相关危害(ASSIST的酒精和THC分量表)的自我报告相比,我们检查了指甲中ETG和羧基-δ-9-THC的敏感性和特异性。然后,我们检查了与指甲和自我报告之间不一致相关的因素。
    结果:几乎三分之二(707/1101)的受访者邮寄指甲剪。那些指甲返回的人不成比例地结婚了,白人种族,年长的,不那么沮丧。在8pg/mg的阈值下,检测危险酒精使用的敏感性为.50,检测酒精相关问题的敏感性为.49。对大麻问题的敏感性只有61。所有措施的特异性均大于0.77。与Audit-C上危险饮酒的阳性指甲/阴性自我报告(即假阳性)相关的因素包括更多的疼痛和未婚;ASSIST上酒精相关问题的假阳性指甲与未婚和非西班牙裔种族有关。ASSIST上THC相关问题的假阳性指甲与非裔美国人有关,西班牙裔,有法律问题。
    结论:在标准截止日期,指甲测量的敏感性低,特异性高.不成比例地提交阳性指甲/阴性自我报告的群体可能具有未被自我报告充分捕获的物质使用模式,由于社会压力,自我报告不准确,或不同的药物代谢。
    BACKGROUND: Nails accumulate the alcohol metabolite, ethyl glucuronide (ETG), and the cannabis metabolite, carboxy- delta-9-THC over 3-6 months. Few studies have examined nail toxicology testing\'s sensitivity and specificity and the agreement between nail testing and self-reported alcohol and marijuana use.
    METHODS: In an ongoing clinical trial, 1101 veterans completed initial telephone questionnaires and were then asked to mail nail clippings for substance use analysis. We examined sensitivity and specificity of ETG and carboxy- delta-9-THC in nails compared to self-report of alcohol use patterns (the AUDIT-C) and substance-related harms (alcohol and THC subscales of the ASSIST). We then examined factors associated with discordance between nails and self-report.
    RESULTS: Almost two-thirds (707/1101) of respondents mailed in nail clippings. Those with returned nails were disproportionately married, white race, older, and less depressed. At a threshold of 8pg/mg, sensitivity was only.50 to detect risky alcohol use and.49 to detect alcohol-related issues. Sensitivity for marijuana issues was only.61. Specificity was greater than.77 for all measures. Factors associated with positive nails/negative self-report (i.e. false positives) for risky alcohol use on the Audit-C included more pain and being unmarried; false positive nails for alcohol-related issues on the ASSIST were associated with being unmarried and non-Hispanic ethnicity. False positive nails for THC-related issues on the ASSIST were associated with being African American, Hispanic, and having had legal issues.
    CONCLUSIONS: At standard cut-offs, nail measures had low sensitivity and higher specificity. The groups who disproportionately submit positive nails/negative self-report could have substance use patterns not adequately captured by self-report, inaccurate self-report due to social pressures, or distinct drug metabolism.
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  • 文章类型: Journal Article
    流动性评分数据可用于估计患病率,发病率,和奶牛群跛行的持续时间。移动性评分通常很少以可变的灵敏度进行,但是这如何影响跛行参数的估计在很大程度上是未知的。我们开发了一个仿真模型,以研究移动性评分的频率和准确性对不同牧群场景的跛行参数估计的影响。在五年的每日时间步长上模拟了具有不同患病率(10、30或50%)和持续时间(分布在中位数第18、36、54、72或108天)的牧群。调查的跛行参数是患病率,持续时间,新病例率,第一次跛行的时间,以及第一年保持声音的可能性。真实参数是从每日数据中计算的,并与复制不同频率时计算的参数进行比较(每周,两周一次,每月,季度),敏感度(60-100%),和移动性评分的特异性(95-100%)。我们的结果表明,当移动性评分的敏感性和特异性<100%时,可能会发生发病率的高估和持续时间的低估。这种效果随着更频繁的评分而增加。在模拟季度流动性评分时,跛行患病率是唯一可以合理估计的参数。这些发现可以帮助告知流动性评分实践和流动性评分数据的解释。
    Mobility scoring data can be used to estimate the prevalence, incidence, and duration of lameness in dairy herds. Mobility scoring is often performed infrequently with variable sensitivity, but how this impacts the estimation of lameness parameters is largely unknown. We developed a simulation model to investigate the impact of the frequency and accuracy of mobility scoring on the estimation of lameness parameters for different herd scenarios. Herds with a varying prevalence (10, 30, or 50%) and duration (distributed around median days 18, 36, 54, 72, or 108) of lameness were simulated at daily time steps for five years. The lameness parameters investigated were prevalence, duration, new case rate, time to first lameness, and probability of remaining sound in the first year. True parameters were calculated from daily data and compared to those calculated when replicating different frequencies (weekly, two-weekly, monthly, quarterly), sensitivities (60-100%), and specificities (95-100%) of mobility scoring. Our results showed that over-estimation of incidence and under-estimation of duration can occur when the sensitivity and specificity of mobility scoring are <100%. This effect increases with more frequent scoring. Lameness prevalence was the only parameter that could be estimated with reasonable accuracy when simulating quarterly mobility scoring. These findings can help inform mobility scoring practices and the interpretation of mobility scoring data.
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  • 文章类型: Journal Article
    背景:逆转录聚合酶链反应(RT-PCR)是用于从呼吸道样品中鉴定COVID-19的主要技术。在几篇文章中已经提出,胸部CT可以为COVID-19提供一种可能的替代诊断工具;然而,没有专业医疗机构建议使用胸部CT作为早期的COVID-19检测方式。这篇文献综述研究了CT扫描作为COVID-19诊断工具的用途。
    方法:使用精确陈述的标准对发表在同行评审期刊上的研究工作进行了全面搜索。搜索仅限于英文出版物,并对使用胸部CT扫描和RT-PCR检测诊断的COVID-19阳性患者进行了研究。对于这篇评论,咨询了四个数据库:这些是Cochrane和ScienceDirect目录,以及EBSCOhost提供的CINAHL和Medline数据库。
    结果:总计,在最初的搜索中发现了285项可能相关的研究。在应用纳入和排除标准后,六项研究仍有待分析。根据纳入的研究,在COVID-19诊断方面,胸部CT扫描显示具有44%至98%的敏感性和25%至96%的特异性。然而,本综述纳入的所有研究均发现了方法学上的局限性.
    结论:RT-PCR仍然是COVID-19的一线诊断技术;虽然胸部CT足以用于有症状的患者,对于COVID-19的初级筛查,它不是一个足够可靠的诊断工具。
    BACKGROUND: Reverse transcription polymerase chain reaction (RT-PCR) is the main technique used to identify COVID-19 from respiratory samples. It has been suggested in several articles that chest CTs could offer a possible alternate diagnostic tool for COVID-19; however, no professional medical body recommends using chest CTs as an early COVID-19 detection modality. This literature review examines the use of CT scans as a diagnostic tool for COVID-19.
    METHODS: A comprehensive search of research works published in peer-reviewed journals was carried out utilizing precisely stated criteria. The search was limited to English-language publications, and studies of COVID-19-positive patients diagnosed using both chest CT scans and RT-PCR tests were sought. For this review, four databases were consulted: these were the Cochrane and ScienceDirect catalogs, and the CINAHL and Medline databases made available by EBSCOhost.
    RESULTS: In total, 285 possibly pertinent studies were found during an initial search. After applying inclusion and exclusion criteria, six studies remained for analysis. According to the included studies, chest CT scans were shown to have a 44 to 98% sensitivity and 25 to 96% specificity in terms of COVID-19 diagnosis. However, methodological limitations were identified in all studies included in this review.
    CONCLUSIONS: RT-PCR is still the suggested first-line diagnostic technique for COVID-19; while chest CT is adequate for use in symptomatic patients, it is not a sufficiently robust diagnostic tool for the primary screening of COVID-19.
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  • 文章类型: Journal Article
    背景:正确诊断结核(TB)淋巴结炎对其治疗和预防至关重要。细针穿刺细胞学(FNAC)是埃塞俄比亚结核病淋巴结炎诊断的主要方法;然而,埃塞俄比亚东部地区尚未评估FNAC的绩效。这项研究旨在评估FNAC和Ziehl-Neelsen(ZN)染色与GeneXpert染色相比在诊断TB淋巴结炎中的表现。
    方法:使用FNAC检查了从291例怀疑患有TB淋巴结炎的患者中收集的细针抽吸(FNA)标本,ZN,和GeneXpert诊断结核淋巴结炎。Gene-Xpert被认为是用于比较的参考标准方法。敏感性,特异性,阳性预测值(PPV),负预测值(NPV),使用SPSS版本25确定κ系数。
    结果:灵敏度,特异性,PPV,ZN诊断结核淋巴结炎的NPV为73.2%,97.4%,96.2%,分别为80.1%。ZN和GeneXpert之间的一致性较差(Kappa=-0.253)。敏感性,特异性,PPV,FNAC的净现值为83.3%,94.8%,93.5%,分别为86.3%。FNAC和GeneXpert之间有适度的一致性(Kappa=0.785)。
    结论:细针穿刺细胞学检查(FNAC)是诊断结核淋巴结炎比ZN更敏感的检查方法。FNAC显示与GeneXpert测定的中等一致性。这项研究建议FNAGeneXpertMTB/RIF测试优先于FNAC来诊断TB淋巴结炎,以避免对涂片阴性的TB淋巴结炎的漏诊。
    BACKGROUND: Proper diagnosis of tuberculosis (TB) lymphadenitis is critical for its treatment and prevention. Fine needle aspirate cytology (FNAC) is the mainstay method for the diagnosis of TB lymphadenitis in Ethiopia; however, the performance of FNAC has not been evaluated in the Eastern Region of Ethiopia. This study aimed to evaluate the performance of FNAC and Ziehl-Neelsen (ZN) staining compared with that of GeneXpert for the diagnosis of TB lymphadenitis.
    METHODS: Fine needle aspiration (FNA) specimens collected from 291 patients suspected of having TB lymphadenitis were examined using FNAC, ZN, and GeneXpert to diagnose TB lymphadenitis. Gene-Xpert was considered the reference standard method for comparison. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and kappa coefficient were determined using SPSS version 25.
    RESULTS: The sensitivity, specificity, PPV, and NPV of ZN for diagnosing TB lymphadenitis were 73.2%, 97.4%, 96.2%, and 80.1% respectively. There was poor agreement between ZN and GeneXpert (Kappa=-0.253). The sensitivity, specificity, PPV, and NPV of FNAC were 83.3%, 94.8%, 93.5%, and 86.3% respectively. There was moderate agreement between the FNAC and GeneXpert (Kappa = 0.785).
    CONCLUSIONS: The fine needle aspiration cytology (FNAC) is a more sensitive test for the diagnosis of TB lymphadenitis than ZN. The FNAC showed a moderate agreement with the GeneXpert assay. This study recommends the FNA GeneXpert MTB/RIF test in preference to FNAC for the diagnosis of TB lymphadenitis to avoid a missed diagnosis of smear-negative TB lymphadenitis.
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  • 文章类型: Journal Article
    球虫寄生虫cyclosporacayetanensis是食源性疾病爆发和每年多次新鲜农产品召回的病原体。这项研究的目的是确定C.cayetanensis18SrRNA和MIT1C基因靶实时定量聚合酶链反应(qPCR)测定的潜在交叉反应物种。评估的环境样本是灌溉池塘水,产生洗涤水,和废水处理污泥来自先前的研究,通过18SrRNA基因靶qPCR对cayetanensis进行qPCR检测。从这些样本中,对18SrRNA基因和线粒体细胞色素c氧化酶亚基III基因(cox3)的较长区域进行了测序。在65个灌溉池塘水样品中,使用C.cayetanensis18SrRNA基因qPCR检测结果为阳性,根据cox3和18SrRNA基因的测序,没有MIT1CqPCR检测或与C.cayetanensis成簇的序列。这些样本的序列聚集在鸟类中发现的球虫序列周围,鱼,爬行动物,和两栖动物宿主。在通过qPCR检测的26个污泥样品中,通过对cox3和18SrRNA基因区域进行测序,可以确认14(54%)含有cayetanensis。在剩余的三个污泥样本中,序列读数与啮齿动物的球虫聚集在一起。这项研究表明,在解释环境样品中的qPCRC.cayetanensis检测数据时应谨慎,可能需要测序步骤进行确认。
    目的:新鲜农产品是环孢素病暴发的主要传播源。因此,必须了解生产生长环境在这种疾病传播中的作用。要做到这一点,必须对环境和灌溉水进行敏感和具体的测试。已确定了环孢霉cayetanensis实时定量聚合酶链反应(qPCR)测定的潜在交叉反应,阻碍了在环境中准确识别这种寄生虫的能力。cox3和18SrRNA基因的扩增子测序显示,最初通过qPCR检测到cayetanensis的所有灌溉池塘水和两个污泥样品最有可能与鸟类脱落的相关球虫生物发生交叉反应,鱼,爬行动物,两栖动物,和啮齿动物。这些结果支持环境样品的单一测试方法可能不足以灵敏和特异地检测cayetanensis。
    The coccidian parasite Cyclospora cayetanensis is the causative agent for foodborne outbreaks of cyclosporiasis disease and multiple annual fresh produce recalls. The aim of this study was to identify potential cross-reacting species for the C. cayetanensis 18S rRNA and MIT1C gene target real-time quantitative polymerase chain reaction (qPCR) assays. The environmental samples evaluated were irrigation pond water, produce wash water, and wastewater treatment sludge from a previous study with qPCR detections of C. cayetanensis by the 18S rRNA gene target qPCR. From these samples, longer regions of the 18S rRNA gene and the mitochondrial cytochrome c oxidase subunit III gene (cox3) were sequenced. Of 65 irrigation pond water samples with positive test results using the C. cayetanensis 18S rRNA gene qPCR assay, none had MIT1C qPCR assay detections or sequences that clustered with C. cayetanensis based on sequencing of the cox3 and 18S rRNA gene. Sequences from these samples clustered around coccidia sequences found in bird, fish, reptile, and amphibian hosts. Of 26 sludge samples showing detections by either qPCR assay, 14 (54%) could be confirmed as containing C. cayetanensis by sequencing of cox3 and 18S rRNA gene regions. In three of the remaining sludge samples, sequenced reads clustered with coccidia from rodents. This study demonstrated that caution should be taken when interpreting qPCR C. cayetanensis detection data in environmental samples and sequencing steps will likely be needed for confirmation.
    OBJECTIVE: Fresh produce is a leading transmission source in cyclosporiasis outbreaks. It is therefore essential to understand the role that produce-growing environments play in the spread of this disease. To accomplish this, sensitive and specific tests for environmental and irrigation waters must be developed. Potential cross-reactions of Cyclospora cayetanensis real-time quantitative polymerase chain reaction (qPCR) assays have been identified, hindering the ability to accurately identify this parasite in the environment. Amplicon sequencing of the cox3 and 18S rRNA genes revealed that all irrigation pond water and two sludge samples that initially detected C. cayetanensis by qPCR were most likely cross-reactions with related coccidian organisms shed from birds, fish, reptiles, amphibians, and rodents. These results support that a single testing method for environmental samples is likely not adequate for sensitive and specific detection of C. cayetanensis.
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