Immunoenzyme Techniques

免疫酶技术
  • 文章类型: Case Reports
    与成人肺炎支原体相关的脑干脑炎(BE)很少见,诊断是具有挑战性的。我们描述了在具有免疫能力的患者中罕见的BE病例。一个43岁的人,其他健康的女人带着高烧和喉咙痛去了我们的急诊室,三天后,她带着昏昏欲睡的精神状态回来了。磁共振成像显示涉及双侧脑桥的双侧大脑区域弥漫性肿胀。通过酶免疫测定(EIA)检测,血清中针对肺炎支原体的免疫球蛋白(Ig)M抗体呈阳性,在医院第10天,针对肺炎支原体的IgM指数水平进一步从1.5增加到2.1。根据肺炎支原体的检测,我们将抗生素方案从万古霉素和头孢曲松改为克拉霉素,我们加了静脉注射免疫球蛋白.一个月后,病人从神经功能缺损中完全康复.进行了脑部磁共振成像的随访,显示病变完全消退。颗粒凝集测定(PA)和EIA都主要用于诊断肺炎支原体。与PA测试相比,EIA测试可能是一个可靠的工具,因为它可以单独测量IgM和IgG抗体.我们通过EIA诊断出与肺炎支原体相关的BE,急性和亚急性配对血清中IgM水平升高。早期使用大环内酯类抗生素治疗可获得良好的结果。
    Brainstem encephalitis (BE) associated with Mycoplasma pneumoniae in adults is rare, and the diagnosis is challenging. We describe an uncommon case of BE in an immunocompetent patient. A 43-year-old, otherwise healthy woman visited our emergency department with high fever and a sore throat, and 3 days later she returned with an altered drowsy mental status. Magnetic resonance imaging displayed diffuse swelling in bilateral cerebral regions involving the bilateral pons. The sera tested positive for the immunoglobulin (Ig) M antibody against M. Pneumoniae as detected by an enzyme immunoassay (EIA), and on hospital day 10, the level of IgM index against M. pneumoniae further increased from 1.5 to 2.1. We changed the antibiotic regimen from vancomycin and ceftriaxone to clarithromycin based on detection of M. pneumoniae, and we added intravenous immunoglobulin. After one month, the patient fully recovered from the neurological deficits. A follow-up brain magnetic resonance imaging was performed, which showed completely resolved lesions. Particle agglutination assay (PA) and EIA are both largely used to diagnose M. pneumoniae. Compared to the PA test, the EIA test could be a reliable tool because it separately measures IgM and IgG antibodies. We diagnosed BE associated with M. pneumoniae through EIA with an increasing level of IgM in the acute and subacute paired sera. Early treatment with macrolide antibiotics resulted in a good outcome.
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  • 文章类型: Journal Article
    目前,诊断COVID-19的唯一可靠检测方法是RT-qPCR。血清学测定已被广泛用于提高感染人群的检测灵敏度。特此,我们报告了一种新的泛IgG多重酶免疫分析(免疫点)方法的性能,用于探索不同的SARS-COV-2血清学结果。
    在罗氏和Snibe平台上,对回收的COVID-19受试者的38份残留血清样本进行了回顾性研究,血清学结果不一致,在新的半自动化泛IgG免疫点酶免疫分析上进行了重新分析,即COVIDOT测试,以找到差异的来源并评估后一种方法。在BlueDiver®仪器上分析所有样品,并且使用D0T®软件通过BlueScan®扫描仪读取所有条带。
    根据我们的数据,受试者样本在免疫点条上对≥2种不同抗原显示特异性IgG反应.在这38个样本中,97.4%的样本显示针对S1+S2抗原的特异性IgG反应,89.5%显示抗RBD抗原,在COVIDOT-TEST试剂盒上86.8%的抗S2抗原反应。在73.7%和65.8%的样本中检测到特异性IgG-S1抗原和IgG-N抗原反应,分别。
    新的半自动化泛IgG免疫点酶免疫测定方法似乎是一种可靠的检测方法,可用于确认可疑的COVID-19血清学筛查结果。
    At present, the only reliable test for COVID-19 diagnosis is RT-qPCR. Serological assays have been widely used to increase the detection sensitivity of infected population. Hereby, we report the performance of a new pan-IgG multiplex Enzyme Immunoassay (immunodot) method for exploration of discrepant SARS-COV-2 serological results.
    A retrospective study on 38 residual serum samples from recovered COVID-19 subjects with discordant serological results on Roche and Snibe platforms, were reanalyzed on a new semi-automated pan-IgG immunodot Enzyme Immunoassay, namely COVIDOT-TEST, in order to find the source of discrepancies and to evaluate the latter method. All samples were analyzed on the BlueDiver® Instrument and all strips were read by the BlueScan® Scanner using Dr DOT® Software.
    Based on our data, subject samples showed specific IgG reactions on ≥  2 different antigens on immunodot strips. Of these 38 samples, 97.4 % of samples showed specific IgG reaction against S1 + S2 antigens, 89.5 % showed against RBD antigen, 86.8 % against S2 antigen reaction on the COVIDOT-TEST kit. Specific IgG-S1 antigen and IgG-N antigen reactions were detected in 73.7 % and 65.8 % of the samples, respectively.
    The new semi-automated pan-IgG immunodot Enzyme Immunoassay method appeared to be a reliable assay to confirm suspicious COVID-19 serological screening results.
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  • 文章类型: Case Reports
    BACKGROUND Few cases of falsely undetectable PSA due to the presence of an inhibitory serum factor have been reported in the world literature. We present a case of falsely low-to-undetectable PSA with data from a serum dilution series, the current literature on biochemical assay interference, and the implications for prostate cancer salvage treatment. CASE REPORT A 63-year-old man was treated with prostatectomy for high-risk prostate cancer and was found to have a rising PSA after approximately 3 years following surgery. He subsequently transferred his care to a different health system and was found to have an undetectable PSA. He was eventually found to have an elevated PSA once again after the particular assay at this institution was changed. He thus received salvage prostate radiotherapy and androgen deprivation therapy. CONCLUSIONS While falsely low PSA results cannot be explained by the presence of serum heterophile antibodies, competitive antibody interference against the immunoassay reagents or anti-PSA antibodies are possible explanations for the results of the dilution experiments performed in this case study. We suggest that unexpected PSA testing results should raise concern for assay interference and warrant further clinical workup.
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  • 文章类型: Journal Article
    BACKGROUND: Clostridioides (formerly Clostridium) difficile infection (CDI) is linked to misuse of antimicrobials. The prevalence of disease varies with difficulties of establishing the diagnosis because of the lack of sensitivity and specificity of laboratory tests. The clinical impact of upgrading CDI testing from routine to molecular based-algorithm is still unclear. The aim of this study is to assess the impact of upgrading CDI testing from routine to molecular based-algorithm on the management of CDI and evaluate the role of antimicrobials on the course of CDI.
    METHODS: This is an observational case-study. A total of 564 patients were included from whom stool samples were tested by enzyme immunoassay (EIA) and Xpert for C. difficile. Data on the number and results of tests ordered, antimicrobial exposure, comorbidities, and treatment with metronidazole or vancomycin were collected. The main outcome measures were C. difficile tests (EIA and Xpert C. difficile Assay) and prevalence of CDI.
    RESULTS: CDI was found in 9 and 10 cases out of 313 and 254 patients tested by the EIA and Xpert C. difficile assay, respectively, giving an overall incidence of 0.03 per 1,000 patient tested. Reduction was noted in the number of tests ordered per patient for presumptive CDI after shifting to the Xpert C. difficile assay which was not statistically significant (p-value 0.2). Also, there was less metronidazole and vancomycin therapy initiated for patients with a negative C. difficile test (p-value 0.2) observed with molecular testing.
    CONCLUSIONS: Xpert C. difficile testing is a supportive tool for diagnosing CDI with rapid turnaround time that is helpful for patient management and initiating effective infection control measures. The clinical accuracy of the assay is still to be determined in the context of low carriage rate in the local patient population.
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  • 文章类型: Case Reports
    Adenoid hypertrophy (AH) is a common disease in children. We describe two cases of AH in children with syphilis enzyme-linked immunosorbent assay (EIA) for false-positive. To our knowledge, there are few reports of false-positive in syphilis EIA in children with AH. Two cases of AH children with syphilis EIA positive samples were confirmed by TPPA and LIA, both showed negative reaction. Therefore, the occurrence of syphilis EIA positive reactions in such diseases should dramatically arouse the attention of docimasters and doctors so as to avoid misdiagnosis caused by false-positive in children with AH.
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  • 文章类型: Case Reports
    背景:评估异位眼眶脑膜瘤的眼科表现和影像学特征,以提高诊断准确性。
    方法:纳入了1999年8月至2017年9月217个月期间入住我们机构的患者的患者数据。患者的眼科表现,影像学特征(CT和MRI),诊断,病理学,治疗方案,并对预后进行回顾性分析。
    结果:确定了6例异位眼眶脑膜瘤患者。首次就诊的平均年龄为33.2岁(范围,7-56岁)。6例患者均表现为眼球突出,上眼睑水肿,和运动障碍,平均病史为20.3个月(范围3-72个月)。光学病变位于鼻上区(3/6,50%),双颞叶外侧区室(1/6,16.7%)和眼眶内侧区室(2/6,33%)。射线照相特征不明确,异质,增强伴有眼外肌粘连(6/6,100%)和钙化(1/6,16.7%)的软组织肿块,不靠近视神经,也不沿硬膜延伸。6例术中完全手术切除,表明所有病变均独立于视神经和蝶骨脊。组织病理学分类主要是脑膜上皮细胞(5/6,83%)。免疫组织化学显示6例EMA和波形蛋白均呈阳性表达,而2例calponin阳性,在嗅球中强烈表达。术后,病变没有引起视力障碍,无复发病例。
    结论:眼眶异位脑膜瘤是一种罕见的肿瘤,不经术后组织病理学检查不易诊断。本报告重点介绍了孤立性眼眶病变的一些显著特征,尤其是额筛骨缝合线周围。观察到伴随的上眼睑水肿和眼睛活动受限与其他眼眶肿瘤不同。在这些情况下,应考虑异位眼眶脑膜瘤的诊断.
    BACKGROUND: To evaluate the ophthalmic manifestations and radiographic features of ectopic orbital meningioma to improve diagnostic accuracy.
    METHODS: Patient data from patients admitted to our institution during a 217-month period from August 1999 to September 2017 were included. Patient ophthalmic manifestations, radiographic features (CT and MRI), diagnosis, pathology, therapeutic regimens, and prognosis were retrospectively reviewed.
    RESULTS: Six patients with ectopic orbital meningioma were identified. The mean age at the first visit was 33.2 years (range, 7-56 years). All six patients displayed manifestations of exophthalmos, upper eyelid oedema, and motility impairment with a mean history of illness of 20.3 months (range 3-72 months). Optical lesions were located in the superonasal extraconal compartment (3/6, 50%), bitemporal extraconal compartment (1/6, 16.7%) and orbital intraconal compartment (2/6, 33%). Radiographic features were ill-defined, heterogeneous, enhancing soft tissue masses with extraocular muscular adhesion (6/6, 100%) and calcification (1/6, 16.7%), not adjacent to the optic nerve and not extending along the dura. Six cases were treated intraoperatively with complete surgical resection, indicating that all lesions were independent of the optic nerve and sphenoid ridge. The histopathologic classification was mostly of meningothelial cells (5/6, 83%). Immunohistochemistry revealed EMA and vimentin to have positive expression in all six cases, while two cases were calponin-positive and strongly expressed in the olfactory bulb. Postoperatively, lesions caused no visual impairment, and there were no cases of recurrence.
    CONCLUSIONS: Ectopic orbital meningiomas are rare tumours that are not easily diagnosed without postoperative histopathology. This report highlights some of the distinguishing features of isolated orbital lesions, especially around the location of frontoethmoidal suture. Accompanying upper eyelid oedema and eye mobility restriction were observed to be dissimilar to other orbital tumours. In these cases, a diagnosis of ectopic orbital meningioma should be considered.
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  • 文章类型: Case Reports
    BACKGROUND: Pulmonary sequestration (PS) is a congenital pulmonary malformation wherein a piece of tissue that ultimately develops into lung tissue is not attached to the pulmonary arterial blood supply, sometimes it is difficult to diagnosis with no specific laboratory tests, discover an abnormal blood supply from aorta by imaging tests is a key step in diagnose.
    METHODS: A 54-year-old male smoker presented with cough, expectoration and blood in the sputum.
    METHODS: Computed tomography (CT) shows lesion in right lung, moderate standard uptake value (SUV) elevation of position-emission tomography/computed tomography (PET/CT) in a part of the lesion, and an increased (1,3)-β-D-glucan assay (G test), Galactomannan enzyme immune-assay (GM test) and tumor marker level, biopsy of lung in different times produced inconclusive results, then finally diagnose of pulmonary sequestration is made by observing an abnormal blood supply from the thoracic aorta and volume change of mass.
    METHODS: The patient refused lower lobectomy which is the main treatment of PS. He was discharged with oral hemostatic and was advised to undergo regular medical checkups.
    RESULTS: The patient has been followed for a year under an outpatient regimen. Symptoms of the cough and expectoration were relieved, however, blood in the sputum remains unchanged.
    CONCLUSIONS: It suggests the need for criteria for a thorough diagnostic work-up. It put emphasis on the importance of considering PS as part of the diagnosis of a lesion in the lung disease and underscore the blood supply of mass. Bronchoscopy or pulmonary lobectomy and follow up of the patient are important for patients diagnosed with pulmonary sequestration.
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  • 文章类型: Journal Article
    Estimation of the effectiveness of rotavirus vaccines via the test-negative control study design has gained popularity over the past few years. In this study design, children with severe diarrhea who test positive for rotavirus infection are considered as cases, while children who test negative serve as controls. We use a simple probability model to evaluate and compare the test-negative control and the traditional case-control designs with respect to the bias of resulting estimates of rotavirus vaccine effectiveness (VE). Comparisons are performed under two scenarios, corresponding to studies performed in high-income and low-income countries. We consider two potential sources of bias: (a) misclassification bias resulting from imperfect sensitivity and specificity of the test used to diagnose rotavirus infection, and (b) selection bias associated with possible effect of rotavirus vaccination on the probability of contracting severe non-rotavirus diarrhea. Our results suggest that both sources of bias may produce VE estimates with substantial bias. Particularly, lack of perfect specificity is associated with severe negative bias. For example, if the specificity of the diagnostic test is 90% then VE estimates from both types of case-control studies may under-estimate the true VE by more than 20%. If the vaccine protects children against non-rotavirus diarrhea then VE estimates from test-negative control studies may be close to zero even though the true VE is 50%. However, the sensitivity and specificity of the enzyme immunoassay test currently used to diagnose rotavirus infections are both over 99%, and there is no solid evidence that the existing rotavirus vaccines affect the rates of non-rotavirus diarrhea. We therefore conclude that the test-negative control study design is a convenient and reliable alternative for estimation of rotavirus VE.
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  • 文章类型: Case Reports
    Blastomyces dermatitidis is a thermally dimorphic fungus that is capable of causing pulmonary and extra-pulmonary disease, including infections of the central nervous system (CNS). Diagnosis of CNS blastomycosis with non-invasive testing can be difficult, and a surgical biopsy may ultimately be required for microbiological and/or histopathological confirmation. A case of B. dermatitidis meningitis is presented where the diagnosis was made by testing cerebrospinal fluid (CSF) using the MVista® Blastomyces Quantitative Antigen Enzyme Immunoassay test. The utility of performing this test on CSF for diagnosis of CNS mass lesions/abscesses caused by B. dermatitidis in the absence of associated meningitis remains unclear. Cross reaction of the Blastomyces antigen test with other dimorphic fungi is a concern, necessitating that positive test results are interpreted in the context of the patient\'s exposure and travel history.
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  • 文章类型: Journal Article
    背景:尿液中的乙基葡糖苷酸(EtG)被认为是最近乙醇消耗或乙醇暴露的标志。叔丁醇主要用作溶剂和中间化学品。像叔戊醇一样,叔丁醇在互联网论坛上讨论作为乙醇替代。我们讨论了通过排泄不同的醇葡糖苷酸(EtG同源物)进行的假阳性免疫EtG筛查,多甲托克科曼病患者尿液中主要是叔丁基葡糖苷酸。
    方法:在BeckmanCoulterAU680分析仪上通过DRIEtG酶免疫测定(ThermoFisherScientificMicrogenics)分析了长期滥用多种物质(包括溶剂)的住院患者的三个连续尿液样本,EtG的内部LC-MS/MS,1-丙基,2-丙基,1-丁基,2-丁基,和叔丁基葡糖苷酸,和自由同类物质甲醇的内部顶空GC-FID,1-丙醇,2-丁酮,2-丁醇,异丁醇,1-丁醇,3-甲基-1-丁醇,2-甲基-1-丁醇,另外对于乙醇,丙酮,2-丙醇,叔丁醇和2-甲基-2-丁醇。
    结果:EtG免疫测定产生两个阳性尿液样本(0.2和0.6mg/L或0.1和0.2mg/g肌酐;截止值0.1mg/L),经LC-MS/MS检测EtG阴性(截止值0.1mg/L),但对叔丁基葡糖苷酸(3.7和27.1mg/L)呈阳性,2-丁基葡糖苷酸(1.1和3.5mg/L),和2-丙基葡糖苷酸(0.1和0.4mg/L)。顶空GC-FID检测叔丁醇(0.97和4.01mg/L),甲醇(0.96和0.62mg/L),2-丁酮(0.84和1.65mg/L),和2-丁醇(0.04和0.09mg/L),但没有乙醇和2-甲基-2-丁醇.
    结论:EtG同系物的交叉反应,怀疑叔丁醇或异丁烷滥用后,主要是叔丁基葡糖苷酸,解释了EtG免疫测定结果的假阳性。未来的研究可以解决尿液中的醇葡糖醛酸苷(EtG同系物)作为(a)对烷烃或其相应的醇代谢产物的生物标志物,以及(b)作为使用“老”-众所周知的醇(如叔丁醇或叔戊醇)的标记容易获得,便宜,有效和“不可检测”的乙醇替代品或“新”的精神活性醇。
    BACKGROUND: Ethyl glucuronide (EtG) in urine is considered a marker of recent ethanol consumption or ethanol exposition. tert-Butanol is primarily used as a solvent and intermediate chemical. Like tert-amyl alcohol, tert-butanol is discussed in Internet forums as ethanol replacement. We discuss false-positive immunological EtG screenings by excretion of different alcohol glucuronides (EtG homologs), mainly tert-butyl glucuronide in urine of a polytoxikomanic in-patient.
    METHODS: Three consecutive urine samples from an in-patient with a long history of multiple substance abuse including solvents were analyzed by DRI EtG enzyme immunoassay (ThermoFisher Scientific Microgenics) on a Beckman Coulter AU680 analyzer, an in-house LC-MS/MS for EtG, 1-propyl, 2-propyl, 1-butyl, 2-butyl, and tert-butyl glucuronide, and an in-house headspace GC-FID of free congener substances methanol, 1-propanol, 2-butanone, 2-butanol, isobutanol, 1-butanol, 3-methyl-1-butanol, 2-methyl-1-butanol, and additionally for ethanol, acetone, 2-propanol, tert-butanol and 2-methyl-2-butanol.
    RESULTS: EtG immunoassay yielded two positive urine samples (0.2 and 0.6mg/L or 0.1 and 0.2mg/g creatinine; cut-off 0.1mg/L) which were tested EtG negative by LC-MS/MS (cut-off 0.1mg/L) but positive for tert-butyl glucuronide (3.7 and 27.1mg/L), 2-butyl glucuronide (1.1 and 3.5mg/L), and 2-propyl glucuronide (0.1 and 0.4mg/L). Headspace GC-FID detected tert-butanol (0.97 and 4.01mg/L), methanol (0.96 and 0.62mg/L), 2-butanone (0.84 and 1.65mg/L), and 2-butanol (0.04 and 0.09mg/L), but no ethanol and no 2-methyl-2-butanol.
    CONCLUSIONS: Cross-reaction of EtG homologs, mainly tert-butyl glucuronide after suspected tert-butanol or isobutane abuse, explains the false-positive EtG immunoassay findings. Future investigations could address the usefulness of alcohol glucuronides (EtG homologs) in urine as (a) biomarkers of an exposition to alkans or their corresponding alcohol metabolites and (b) as markers for using \"old\"-well known alcohols like tert-butanol or tert-amyl alcohol as easy to obtain, cheap, potent and \"undetectable\" ethanol replacements or \"New\" Psychoactive Alcohols.
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