Screening of antinuclear antibodies: comparison between enzyme immunoassay based on nuclear homogenates, purified or recombinant antigens and immunofluorescence assay

2004 ◽  
Vol 42 (10) ◽  
Author(s):  
Sergio Bernardini ◽  
Maria Infantino ◽  
Lorenza Bellincampi ◽  
Marzia Nuccetelli ◽  
Antonella Afeltra ◽  
...  
Keyword(s):  
Screening Test ◽  
Antinuclear Antibody ◽  
Large Scale ◽  
Antinuclear Antibodies ◽  
Immunofluorescence Assay ◽  
Choice Test ◽  
Recombinant Antigens ◽  
Specificity And Sensitivity ◽  
First Choice ◽  
Κ Statistic

AbstractCurrent clinical practice considers antinuclear antibody (ANA) testing as a screening test; this has a major impact on laboratory work with a growing volume of analyses that need to be performed rapidly, to maintain good specificity and sensitivity. Ongoing discussions have been raised in order to identify the best technology to use in ANA screening, taking into account both clinical and economical implications. The aim of our study was to compare three different enzyme immunoassays (EIA) with immunofluorescence (IF) assay in order to identify which test is better for use as a screening test. The study was performed on 473 sera and the three different EIA tests were based on nuclear homogenates from HeLa cells, purified antigens from HEp-2 cells and recombinant antigens, respectively. The concordance between EIA-ANA and IF-ANA techniques, determined by the κ statistic, was acceptable, but not complete, and discrepancies between both EIA-positive/IF-negative samples and IF-positive/EIA-negative were found. Both methods show interesting diagnostic abilities, however, the IF-ANA assay seems to be the first choice test in a well-standardized immunofluorescence laboratory with experienced microscopists, whereas the EIA test might be useful especially in large-scale ANA screening.

Antinuclear Antibodies Testing Method Variability: A Survey of Participants in the College of American Pathologists’ Proficiency Testing Program

2020 ◽  
Vol 47 (12) ◽  
pp. 1768-1773
Author(s):  
Stanley J. Naides ◽  
Jonathan R. Genzen ◽  
Gyorgy Abel ◽  
Christine Bashleben ◽  
M. Qasim Ansari
Keyword(s):  
Antinuclear Antibody ◽  
Antinuclear Antibodies ◽  
Screening Method ◽  
Immunofluorescence Assay ◽  
Test Target ◽  
Testing Method ◽  
Reporting Practice ◽  
Cell Substrate ◽  
Target Performance ◽  
Automated Platform

ObjectiveThis study was conducted to determine the spectrum of laboratory practices in antinuclear antibody (ANA) test target, performance, and result reporting.MethodsA questionnaire on ANA testing was distributed by the Diagnostic Immunology and Flow Cytometry Committee of the College of American Pathologists (CAP) to laboratories participating in the 2016 CAP ANA proficiency survey.ResultsOf 5847 survey kits distributed, 1206 (21%) responded. ANA screening method varied: 55% indirect immunofluorescence assay, 21% ELISA, 12% multibead immunoassay, and 18% other methods. The name of the test indicated the method used in only 32% of laboratories; only 39% stated the method used on the report. Of 644 laboratories, 80% used HEp-2 cell substrate, 18% HEp-2000 (HEp-2 cell line engineered to overexpress SSA antigen, Ro60), and 2% other. Slides were prepared manually (67%) or on an automated platform (33%) and examined by direct microscopy (84%) or images captured by an automated platform (16%). Only 50% reported a positive result at the customary 1:40 dilution. Titer was reported to endpoint routinely by 43%, only upon request by 23%, or never by 35%. Of the laboratories, 8% did not report dual patterns. Of those reporting multiple patterns, 23% did not report a titer with each pattern.ConclusionANA methodology and practice, and test naming and reporting varies significantly between laboratories. Lack of uniformity in testing and reporting practice and lack of transparency in communicating the testing method may misdirect clinicians in their management of patients.

scholarly journals A computer program that periodically monitors the ability to interpret the antinuclear antibody test

1996 ◽  
Vol 42 (5) ◽  
pp. 836-840 ◽  
Author(s):  
M L Astion ◽  
M H Wener ◽  
K Hutchinson ◽  
G B Olsen ◽  
A R Orkand ◽  
...  
Keyword(s):  
Antinuclear Antibody ◽  
Antinuclear Antibodies ◽  
Antibody Test ◽  
Computer Programs ◽  
Immunofluorescence Assay ◽  
Image Features ◽  
Test Results ◽  
Medical Technologists ◽  
Logical Extension ◽  
Gain Access

Abstract Our laboratory has been developing computer programs that help medical technologists improve their performance of the microscope-based immunofluorescence assay for antinuclear antibodies (ANA). This image-based laboratory test has been associated with poor reproducibility. We have previously described our first program, ANA-Tutor, which systematically teaches the ANA test by using approximately 150 processed digital images of ANA test results. The program we describe here, Pattern Plus Auditor, is a logical extension to ANA-Tutor. Pattern Plus Auditor tests the ability of laboratory personnel to interpret the ANA test, and tracks individual and laboratory performance over time. The program consists of image-based questions that test a variety of ANA staining patterns, including homogeneous, speckled, centromere, nucleolar, mixed patterns, and rare patterns. For each question, the program provides correct answers with explanations and color overlays that highlight key image features. By entering the proper password, users gain access to exam results for individuals and for the laboratory as a whole. Results are available for the current exam, any previous exam, or cumulatively on all exams to date. Intralaboratory testing with computer programs such as Pattern Plus Auditor might be a useful part of quality-assurance procedures for many image-based laboratory tests.

Methods for the Quantification of Salivary Cortisol and of a-amylase in Biosensors and Portable Devices

2018 ◽  
Vol 68 (12) ◽  
pp. 2857-2859
Author(s):  
Cristina Mihaela Ghiciuc ◽  
Andreea Silvana Szalontay ◽  
Luminita Radulescu ◽  
Sebastian Cozma ◽  
Catalina Elena Lupusoru ◽  
...  
Keyword(s):  
Real Time ◽  
Medical Practice ◽  
Salivary Cortisol ◽  
Clinical Studies ◽  
Large Scale ◽  
Psychological Research ◽  
Portable Devices ◽  
Salivary Amylase ◽  
Specificity And Sensitivity

There is an increasing interest in the analysis of salivary biomarkers for medical practice. The objective of this article was to identify the specificity and sensitivity of quantification methods used in biosensors or portable devices for the determination of salivary cortisol and salivary a-amylase. There are no biosensors and portable devices for salivary amylase and cortisol that are used on a large scale in clinical studies. These devices would be useful in assessing more real-time psychological research in the future.

scholarly journals Assessment of antinuclear antibodies by indirect immunofluorescence assay: report from a survey by the American Association of Medical Laboratory Immunologists

2020 ◽  
Vol 58 (9) ◽  
pp. 1489-1497 ◽  
Author(s):  
Lisa K. Peterson ◽  
Anne E. Tebo ◽  
Mark H. Wener ◽  
Susan S. Copple ◽  
Marvin J. Fritzler
Keyword(s):  
Indirect Immunofluorescence ◽  
Antinuclear Antibodies ◽  
Current Practice ◽  
Clinical Laboratory ◽  
Immunofluorescence Assay ◽  
Indirect Immunofluorescence Assay ◽  
Medical Laboratory ◽  
International Consensus ◽  
Clinical Laboratories ◽  
Reading Interpretation

AbstractBackgroundThe indirect immunofluorescence assay (IFA) using HEp-2 cell substrates is the preferred method by some for detecting antinuclear antibodies (ANA) as it demonstrates a number of characteristic staining patterns that reflect the cellular components bound as well as semi-quantitative results. Lack of harmonized nomenclature for HEp-2 IFA patterns, subjectivity in interpretation and variability in the number of patterns reported by different laboratories pose significant harmonization challenges. The main objectives of this study were to assess current practice in laboratory assessment of HEp-2 IFA, identify gaps and define strategies to improve reading, interpretation and reporting.MethodsWe developed and administered a 24-item survey based on four domains: educational and professional background of participants, current practice of HEp-2 IFA testing and training, gap assessment and the perceived value of International Consensus on Antinuclear Antibody Patterns (ICAP) and other factors in HEp-2 IFA assessment. The Association of Medical Laboratory Immunologists (AMLI) and American Society for Clinical Pathology administered the survey from April 1 to June 30, 2018, to members involved in ANA testing. This report summarizes the survey results and discussion from a dry workshop held during the 2019 AMLI annual meeting.ResultsOne hundred and seventy-nine (n = 179) responses were obtained where a significant number were clinical laboratory scientists (46%), laboratory directors (24%), supervisors (13%) or others (17%). A majority of respondents agreed on the need to standardize nomenclature and reporting of HEp-2 IFA results. About 55% were aware of the ICAP initiative; however, among those aware, a significant majority thought its guidance on HEp-2 IFA nomenclature and reporting is of value to clinical laboratories. To improve ICAP awareness and further enhance HEp-2 IFA assessment, increased collaboration between ICAP and the clinical laboratory community was suggested with emphasis on education and availability of reference materials.ConclusionsBased on these suggestions, future efforts to optimize HEp-2 IFA reading, interpretation and reporting would benefit from more hands-on training of laboratory personnel as well as continuous collaboration between professional organizations, in vitro diagnostic manufacturers and clinical laboratories.

scholarly journals Bayesian estimation of the seroprevalence of antibodies to SARS-CoV-2

JAMIA Open ◽  
2020 ◽  
Author(s):  
Qunfeng Dong ◽  
Xiang Gao
Keyword(s):  
Probability Distribution ◽  
Large Scale ◽  
Prior Probability ◽  
Probability Distributions ◽  
Single Point ◽  
Point Estimates ◽  
Specificity And Sensitivity ◽  
Large Scale Dataset ◽  
The Us ◽  
Antibody Tests

Abstract Accurate estimations of the seroprevalence of antibodies to severe acute respiratory syndrome coronavirus 2 need to properly consider the specificity and sensitivity of the antibody tests. In addition, prior knowledge of the extent of viral infection in a population may also be important for adjusting the estimation of seroprevalence. For this purpose, we have developed a Bayesian approach that can incorporate the variabilities of specificity and sensitivity of the antibody tests, as well as the prior probability distribution of seroprevalence. We have demonstrated the utility of our approach by applying it to a recently published large-scale dataset from the US CDC, with our results providing entire probability distributions of seroprevalence instead of single-point estimates. Our Bayesian code is freely available at https://github.com/qunfengdong/AntibodyTest.

scholarly journals SARS-CoV-2 sample-to-answer nucleic acid testing in a tertiary care emergency department: evaluation and utility

2020 ◽  
Author(s):  
Pia Jokela ◽  
Anu E Jääskeläinen ◽  
Hanna Jarva ◽  
Tanja Holma ◽  
Maarit J Ahava ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Emergency Departments ◽  
Large Scale ◽  
Tertiary Care ◽  
Reference Method ◽  
Preliminary Evaluation ◽  
Specificity And Sensitivity ◽  
Department Evaluation ◽  
Test Result ◽  
Respiratory Specimens

AbstractRapid sample-to-answer tests for detection of SARS-CoV-2 are emerging and data on their relative performance is urgently needed. We evaluated the analytical performance of two rapid nucleic acid tests, Cepheid Xpert® Xpress SARS-CoV-2 and Mobidiag Novodiag® Covid-19, in comparison to a combination reference of three large-scale PCR tests. Moreover, utility of the Novodiag® test in tertiary care emergency departments was assessed. In the preliminary evaluation, analysis of 90 respiratory samples resulted in 100% specificity and sensitivity for Xpert®, whereas analysis of 107 samples resulted in 93.4% sensitivity and 100% specificity for Novodiag®. Rapid SARS-CoV-2 testing with Novodiag® was made available for four tertiary care emergency departments in Helsinki, Finland between 18 and 31 May, coinciding with a rapidly declining epidemic phase. Altogether 361 respiratory specimens, together with relevant clinical data, were analyzed with Novodiag® and reference tests: 355/361 of the specimens were negative with both methods, and 1/361 was positive in Novodiag® and negative by the reference method. Of the 5 remaining specimens, two were negative with Novodiag®, but positive with the reference method with late Ct values. On average, a test result using Novodiag® was available nearly 8 hours earlier than that obtained with the large-scale PCR tests. While the performance of novel sample-to-answer PCR tests need to be carefully evaluated, they may provide timely and reliable results in detection of SARS-CoV-2 and thus facilitate patient management including effective cohorting.

scholarly journals Comparative study of effectiveness of pap smear versus visual inspection with acetic acid for mass screening of lesions of cervix

2018 ◽  
Vol 6 (6) ◽  
pp. 2042
Author(s):  
Mercy Mrudula Dasari ◽  
Venkatalakshmi Anem ◽  
Sirisha Gunta ◽  
Satish Kumar Seeram ◽  
Bhagyalakshmi Atla
Keyword(s):  
Acetic Acid ◽  
Epithelial Cell ◽  
Screening Test ◽  
Visual Inspection ◽  
Pap Smear ◽  
Cervical Lesions ◽  
Papanicolaou Test ◽  
Promising Alternative ◽  
Screening Programs ◽  
Specificity And Sensitivity

Background: Cervical cancer is a leading cause of morbidity and mortality among women worldwide and more over in the developing countries, so there is a need to develop screening test with high specificity and sensitivity. Aim of the study was to evaluate the effectiveness of Papanicolaou (PAP) smear versus visual inspection acetic acid (VIA) for screening cervical lesions in patients and to determine and compare their sensitivity and specificity.Methods: The present study is a hospital based prospective study for a period of two years at the department of pathology from August 2014 to July 2016 consisting of 500 patients attending gynaecology outpatient clinic. Papanicolaou (Pap) smear tests and visual inspection acetic acid were employed along with complete clinical history record. The results of VIA were correlated with that of pap smear on the basis of sensitivity, specificity and positive and negative predictive value.Results: Out of 500 cases, most common age group was 21 to 40 years of age consisting of 305 cases - 61%. VIA was positive in 156 cases-31.2%, PAP smear positive for epithelial cell abnormalities were 60 cases-12%. VIA showed higher sensitivity (52.38%) compared to Pap smear (40%) whereas Pap smear showed higher specificity (93.2%) compared to VIA (92.4%).Conclusions: Papanicolaou test is a better screening test for epithelial cell abnormality than VIA. However, in countries with low resource settings where cytology-based screening programs are not available, VIA is a promising alternative.

scholarly journals Elevated Concentrations of Soluble Fas and FasL in Multiple Sclerosis Patients with Antinuclear Antibodies

2020 ◽  
Vol 9 (12) ◽  
pp. 3845
Author(s):  
Josip Sremec ◽  
Sanja Tomasović ◽  
Nada Tomić Sremec ◽  
Alan Šućur ◽  
Jelena Košćak Lukač ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Antinuclear Antibodies ◽  
Disease Duration ◽  
Immunofluorescence Assay ◽  
Peripheral Lymphocyte ◽  
Effective Control ◽  
Lymphocyte Apoptosis ◽  
Soluble Fas ◽  
Potential Link ◽  
Multiple Sclerosis Patients

Antinuclear antibodies (ANA) are currently considered as an epiphenomenon of apoptotic processes, possibly in control of autoreactivity in patients with multiple sclerosis (MS). Apoptosis of reactive lymphocytes by the Fas/FasL system is described as an effective control mechanism for autoreactivity in MS. We aimed to provide a context to the potential link between ANA and peripheral lymphocyte apoptosis in MS. The presence of ANA was detected in sera by immunofluorescence assay, and concentrations of sFas and sFasL were determined in the sera of 44 and cerebrospinal fluid (CSF) of 11 relapsing-remitting (RR) MS patients using cytometric bead-based array, and their association with the disease characteristics was determined. ANA were detected in the sera of 43.2% of RRMS patients, and their frequency was the highest in patients with disease duration of less than one year (88,89%). In addition, the number of experienced relapses was lower in ANA-positive patients. Concentrations of sFasL were inversely associated with patients’ expanded disability status scale (EDSS) scores. Low concentrations of both soluble factors strongly discriminated patients with moderate to severe disability, from patients with mild or absent disability only in a group of patients with prolonged disease duration (>10 years). Both soluble mediators were significantly higher in ANA-positive patients. FasL concentrations were inversely associated with the number of relapses. There is a potential link between the presence of ANA and peripheral lymphocyte apoptosis mediated by Fas/FasL system in MS, whose precise role and significance needs to be determined by future mechanistic studies.

scholarly journals SARS-CoV-2 S1 and N-based serological assays reveal rapid seroconversion and induction of specific antibody response in COVID-19 patients

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Abdullah Algaissi ◽  
Mohamed A. Alfaleh ◽  
Sharif Hala ◽  
Turki S. Abujamel ◽  
Sawsan S. Alamri ◽  
...  
Keyword(s):  
Antibody Response ◽  
Specific Antibody ◽  
Large Scale ◽  
Disease Onset ◽  
High Specificity ◽  
Epidemiological Studies ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specificity And Sensitivity ◽  
Human Coronaviruses

Abstract As the Coronavirus Disease 2019 (COVID-19), which is caused by the novel SARS-CoV-2, continues to spread rapidly around the world, there is a need for well validated serological assays that allow the detection of viral specific antibody responses in COVID-19 patients or recovered individuals. In this study, we established and used multiple indirect Enzyme Linked Immunosorbent Assay (ELISA)-based serological assays to study the antibody response in COVID-19 patients. In order to validate the assays we determined the cut off values, sensitivity and specificity of the assays using sera collected from pre-pandemic healthy controls, COVID-19 patients at different time points after disease-onset, and seropositive sera to other human coronaviruses (CoVs). The developed SARS-CoV-2 S1 subunit of the spike glycoprotein and nucleocapsid (N)-based ELISAs not only showed high specificity and sensitivity but also did not show any cross-reactivity with other CoVs. We also show that all RT-PCR confirmed COVID-19 patients tested in our study developed both virus specific IgM and IgG antibodies as early as week one after disease onset. Our data also suggest that the inclusion of both S1 and N in serological testing would capture as many potential SARS-CoV-2 positive cases as possible than using any of them alone. This is specifically important for tracing contacts and cases and conducting large-scale epidemiological studies to understand the true extent of virus spread in populations.

scholarly journals Laboratory Confirmation of Lyme Disease

1991 ◽  
Vol 2 (2) ◽  
pp. 64-69 ◽  
Author(s):  
Tom G Schwan ◽  
Warren J Simpson ◽  
Patricia A Rosa
Keyword(s):  
Lyme Disease ◽  
Immunofluorescence Assay ◽  
Detection Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Tests ◽  
Specificity And Sensitivity ◽  
Laboratory Confirmation ◽  
Staining Methods ◽  
Alternative Detection ◽  
Species Specific

Lyme disease can be confirmed in the laboratory by isolation or detection of its causative agent, a tick-borne spirocheteBorrelia burgdorferi,or by a diagnostic change in the titre of antibodies specific to the agent.B burgdorferican be isolated and cultivated in Barbour-Stoenner-Kelly II medium. It can be detected by light microscopy in tissue sections or, rarely, in blood smears using various staining methods. There is interest in the development of alternative detection methods, including identification of specific antigens ofB burgdorferiin the urine of suspected cases and demonstration of the presence of species-specific DNA using polymerase chain reaction assays. Currently, serological tests (indirect immunofluorescence assay, enzyme-linked immunosorbent assay and Western immunoblot) are the most practical and available methods for confirming Lyme disease. The quest to improve the specificity and sensitivity of serological tests – for example, through the use of specific recombinant antigens – continues.

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