Development and comparative evaluation of immunoblot assays for detecting autoantibodies to Scl 70 and Jo 1 antigens in serum

1990 ◽  
Vol 36 (12) ◽  
pp. 2053-2056 ◽  
Author(s):  
T Fonong ◽  
S M Evans ◽  
H A Homburger
Keyword(s):  
Comparative Evaluation ◽  
Clinical Laboratory ◽  
Connective Tissue Diseases ◽  
Positive Control ◽  
Analytical Sensitivity ◽  
Healthy Controls ◽  
Clinical Use ◽  
Immunoblot Assay ◽  
Negative Results ◽  
Positive Results

Abstract We developed rapid 24-h immunoblot assays for detecting autoantibodies to Scl 70 and Jo 1 antigens in serum. In comparative studies, we evaluated the analytical sensitivity of the immunoblot assays and commercial immunodiffusion assays for anti-Scl 70 and anti-Jo 1 autoantibodies with the use of positive control sera, and compared the frequencies of positive and negative results in a group of 116 sera, including specimens from 34 healthy controls and 82 patients with various connective-tissue diseases. The immunoblot assays were greater than 100-fold more sensitive than immunodiffusion for detecting both autoantibodies. Despite greater analytical sensitivity, there were no false-positive results by the immunoblot assay for anti-Scl 70 or anti-Jo 1 autoantibodies in sera from either the controls or the patients. The diagnostic sensitivity of the immunoblot assay for anti-Scl 70 autoantibodies in patients with scleroderma was greater than that of the immunodiffusion assay, 70% vs 20%, and was equivalent in patients with polymyositis, 43%. We conclude that rapid immunoblot assays for anti-Scl 70 and anti-Jo 1 autoantibodies are superior to immunodiffusion assays for clinical use and are suitable for routine use in the clinical laboratory.

scholarly journals Usefulness of real-time PCR assay targeting lipL32 gene for diagnosis of human leptospirosis in Uruguay

2013 ◽  
Vol 7 (12) ◽  
pp. 941-945 ◽  
Author(s):  
Sabina González ◽  
Juan Pablo Geymonat ◽  
Elba Hernández ◽  
Juan Martín Marqués ◽  
Felipe Schelotto ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Dna Amplification ◽  
Diagnostic Sensitivity ◽  
Analytical Sensitivity ◽  
Serum Samples ◽  
Initial Management ◽  
Negative Results ◽  
Acute Infections ◽  
Positive Results

Introduction: Assays based on DNA amplification can provide information that contributes to the initial management of patients with leptospirosis. However, these have not been adopted in Uruguay. Our aim was to evaluate the performance of the lipL32 real-time PCR (qPCR) for diagnosis of leptospirosis. Methodology: We analyzed by microscopic agglutination test (MAT) and lipL32 qPCR serum samples from 183 patients with suspected leptospirosis. To establish the analytical sensitivity of the qPCR, experimentally spiked samples with known amounts of Leptospira interrogans were analyzed. Results: The analytical sensitivity of the qPCR was 102 leptospires/mL. In 98 patients MAT results were negative meanwhile 85 showed positive reactions, revealing acute infections. Twenty six acute-phase sera of these 85 patients showed a positive signal by qPCR (diagnostic sensitivity 30%). In these patients the average time between onset of symptoms and collection of the first sample was 8 days. In patients with negative results for qPCR and positive MAT results (n=59) the average interval between onset of symptoms and collection of the first sample was 13 days. The qPCR did not yield false positive results. Conclusions: The qPCR had a lower diagnostic sensitivity than MAT and a higher cost. However, it allowed to make an early diagnosis in 26 patients. In patients with confirmed acute infections and negative results by qPCR, more than 8 days had elapsed between the onset of the illness and extraction of the first serum sample. Our data support that the qPCR from sera have clinical utility within the first week of illness.

scholarly journals Immunoblot Assay Using Recombinant Antigens as a Supplemental Test To Confirm the Presence of Antibodies to Trypanosoma cruzi

2007 ◽  
Vol 14 (4) ◽  
pp. 355-361 ◽  
Author(s):  
Kevin Y. Cheng ◽  
Chi-Deu Chang ◽  
Vince A. Salbilla ◽  
Louis V. Kirchhoff ◽  
David A. Leiby ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
False Positive ◽  
Test Results ◽  
Serological Tests ◽  
Assay Sensitivity ◽  
Immunoblot Assay ◽  
Recombinant Antigens ◽  
Persistent Problem ◽  
Negative Results ◽  
Positive Results

ABSTRACT The diagnosis of chronic Chagas' disease is generally made by detecting antibodies to Trypanosoma cruzi. Most conventional serological tests are based on lysates of whole parasites or semipurified antigen fractions from T. cruzi epimastigotes grown in culture. The occurrence of inconclusive and false-positive results has been a persistent problem with the conventional assays, and there is no universally accepted gold standard for confirmation of positive test results. We describe here an immunoblot assay for detecting antibodies to T. cruzi in which four chimeric recombinant antigens (rAgs), designated FP3, FP6, FP10, and TcF, are used as target antigens. Each of these rAgs is composed of several antigenically distinct regions and includes repetitive as well as nonrepetitive sequences. Each rAg is coated as a discrete line on a nitrocellulose strip. Assay sensitivity was assessed by testing 345 specimens known to be positive for antibodies to T. cruzi. All 345 of these samples showed two to four reactive test bands in addition to the three on-board control bands that are on each strip. Assay specificity was determined by testing 500 specimens from random U.S. blood donors, all of which gave negative results. Based on the results obtained in this study, we propose the following scheme for interpretation of test results: (i) no bands or a single test band = a negative result; (ii) two or more test bands with at least one band showing intensity of 1+ or higher = a positive result; and (iii) multiple faint test bands (±) = indeterminate result. Based on this scheme, the prototype immunoblot assay showed sensitivity of 100% (n = 345) and specificity of 100% (n = 500). Additionally, all 269 potentially cross-reacting and T. cruzi antibody-negative specimens tested negative in our immunoblot assay. The rAg-based immunoblot assay has potential as a supplemental test for confirming the presence of antibodies to T. cruzi in blood specimens and for identifying false-positive results obtained with other assays.

Highly sensitive qualitative methods for serum choriogonadotropin (hCG): clinical specificity studies.

1987 ◽  
Vol 33 (5) ◽  
pp. 677-681 ◽  
Author(s):  
Z L Bandi ◽  
I Schoen ◽  
M DeLara
Keyword(s):  
False Positive ◽  
Quantitative Methods ◽  
Analytical Sensitivity ◽  
Negative Results ◽  
Beta Hcg ◽  
Highly Sensitive ◽  
Menopausal Women ◽  
Pregnancy Testing ◽  
Post Menopausal ◽  
Positive Results

Abstract We screened six highly sensitive kits, designed for serum pregnancy tests, for false-positive results. The two best were then evaluated more extensively. The "BETA-hCG MAIA-clone" (I) and the "TANDEM ICON" (II) kits gave only negative results for 100 sera from men at 5, 10, and 25 int. units/L (1st IRP). Of 100 serum specimens from post-menopausal women three and 10 were hCG positive by the II and the I reagents, respectively, but only at the 5 int. units (1st IRP) of hCG per liter level of sensitivity. At 10 and 25 int. units/L, all specimens were negative by both tests. The manufacturers of these kits recommend pregnancy testing only at the 25 int. units/L level of analytical sensitivity. By quantitative methods, hCG concentrations in the II positive samples ranged from 6 to 20 int. units/L (1st IRP) and lutropin concentrations were between 130 to greater than 150 int. units/L. The medical records of the corresponding patients did not support the presence of trophoblast or any other hCG-secreting tissues. During 15 months of routine use of the II reagents at an analytical sensitivity of 25 int. units/L (1st IRP) hCG for pregnancy testing (greater than 4000 serum specimens from pre-menopausal women), the staff has not reported to us any suspected false-positive findings.

Analytical Validation of qPCR-Based Multivariate Index Assays in a Clinical Laboratory: Practical Challenges and Limitations

2018 ◽  
Vol 3 (2) ◽  
pp. 267-281
Author(s):  
Cheryl L Sesler ◽  
Elena V Grigorenko
Keyword(s):  
Clinical Laboratory ◽  
Disease Status ◽  
Analytical Performance ◽  
Analytical Sensitivity ◽  
Clinical Use ◽  
Analytical Validation ◽  
Clinical Laboratories ◽  
Practical Strategy ◽  
Regulatory Guidelines ◽  
Performance Specifications

Abstract Background Multivariate index assays (MIAs) to evaluate disease status and/or therapeutic efficacy are increasingly being used in clinical laboratories as laboratory-developed tests (LDTs). Before clinical use, diagnostic and analytical performance specifications of LDTs must be established. Several regulatory guidelines have been published that address specific components of validation procedures, but the interpretation for the analytical validation of MIAs is ambiguous and creates confusion when implementing a novel MIA in the clinical laboratory. Content CLSI guidelines and published methods were evaluated to develop a validation strategy to establish analytical sensitivity, precision, specificity, and stability for qPCR-based MIAs. Limitations and challenges identified while evaluating guidelines and literature and implementing this strategy are discussed in this review, including sample sourcing and integrity, laboratory contamination, and sample throughput. Due to the diversity of qPCR-based MIAs, we discuss additional considerations for researchers intending to transfer MIAs to a clinical laboratory. Summary A practical strategy to assess the analytical performance characteristics for validation of qPCR-based MIAs was developed and tested before diagnostic clinical use. Several important limitations, challenges, and considerations were identified during development of the analytical validation procedures that are not addressed in regulatory guidelines or published literature. The described strategy can provide insight for future developers of MIAs and clinical laboratories implementing MIAs as LDTs.

Critical Evaluation of Impedance Phlebography for the Diagnosis of Deep Vein Thrombosis

1974 ◽  
Vol 31 (02) ◽  
pp. 273-278
Author(s):  
Kenneth K Wu ◽  
John C Hoak ◽  
Robert W Barnes ◽  
Stuart L Frankel
Keyword(s):  
Deep Vein Thrombosis ◽  
False Positive ◽  
False Negative ◽  
Critical Evaluation ◽  
Original Method ◽  
Vein Thrombosis ◽  
Negative Results ◽  
False Negative Results ◽  
Deep Vein ◽  
Positive Results

SummaryIn order to evaluate its daily variability and reliability, impedance phlebography was performed daily or on alternate days on 61 patients with deep vein thrombosis, of whom 47 also had 125I-fibrinogen uptake tests and 22 had radiographic venography. The results showed that impedance phlebography was highly variable and poorly reliable. False positive results were noted in 8 limbs (18%) and false negative results in 3 limbs (7%). Despite its being simple, rapid and noninvasive, its clinical usefulness is doubtful when performed according to the original method.

PET/CT WITH 68GA-PSMA-11 IN DETECTION OF PRIMARY TUMOR AND RECURRENCE OF PROSTATE CANCER

2018 ◽  
Vol 64 (6) ◽  
pp. 799-804
Author(s):  
Darya Ryzhkova ◽  
M. Poyda
Keyword(s):  
Prostate Cancer ◽  
Primary Tumor ◽  
Biochemical Recurrence ◽  
Whole Body ◽  
Negative Results ◽  
Primary Prostate Cancer ◽  
Pet Ct ◽  
The Relationship ◽  
Positive Results ◽  
T Category

Purpose: To study the diagnostic value of PET-CT with 68Ga-PSMA-11 in the diagnosis of a primary prostate cancer, preoperative staging, and the detection of recurrence of prostate cancer (PCa). Methods: 28 patients aged 64.7 ± 8.74 years were included. 10 patients primary prostate cancer, and 18 patients with biochemical recurrence of the disease after radical treatment were examined. All patients underwent PET-CT with 68Ga-PSMA-11 according the whole body protocol. Interpretation of images was performed visually and quantitatively by calculation of SUL max. Results: High focal or diffuse 68Ga-PSMA-11 uptake was found in prostate parenchyma in patients with primary prostate cancer. Additionally metastases in regional lymph nodes were diagnosed in 4 patients and bone metastases were found in one patient. The correlation between 68Ga-PSMA-11 uptake level and Gleason index in the primary tumor (R Spearmen = 0.25, p = 0.57) was not observed. PET-positive results were obtained in 14 patients and PET-negative results in 4 patients with biochemical recurrence of PCa. The relationship between the frequency of PET-positive results and Gleason index was not revealed (R Spearmen = 0.2, p = 0.39). We found a weak but significant correlation between the frequency of PET-positive results and the prostate tumor stage according to the T category (R Spearmen = 0.49, p = 0.049). In patients with low values of PSA (less than 1.0 ng/ml) in 4 out of 9 cases, PET-negative results were obtained. In patients with PSA level more than 1.0 ng/ml PET-positive results were obtained in all cases. Conclusions: PET/CT with 68Ga-PSMA-11 allows to diagnose the primary prostate cancer, to establish the stage of the disease in categories N and M, and also to determine the localization and dissemination of the tumor in patients with biochemical recurrence of prostate cancer. The relationship between 68Ga-PSMA-11 uptake in primary tumor and Gleason index was not found. The probability of obtaining PET-positive results in cases of biochemical recurrence is affected by a PSA level above 1 ng/ml and a high stage of the disease according to the T category (T3-T4).

scholarly journals Diagnostic Performance of Ankle-Brachial Pressure Index in Lower Extremity Arterial Disease

2021 ◽  
Vol 07 (03) ◽  
pp. e132-e137
Author(s):  
Mohammed Alagha ◽  
Thomas M. Aherne ◽  
Ahmed Hassanin ◽  
Adeel S. Zafar ◽  
Doireann P. Joyce ◽  
...  
Keyword(s):  
Lower Extremity ◽  
Sensitivity And Specificity ◽  
False Negative ◽  
Arterial Disease ◽  
Initial Assessment ◽  
Reference Standard ◽  
Lower Extremity Arterial Disease ◽  
Negative Results ◽  
Positive Results ◽  
Arterial Imaging

Abstract Introduction Ankle-brachial pressure indices (ABIs) continue to form the basis of diagnostics for lower extremity arterial disease (LEAD). However, there remains a paucity of data to support its accuracy. This study aims to evaluate its diagnostic sensitivity and specificity using established arterial-imaging modalities as a benchmark. Methods In this retrospective study, a regional, prospectively maintained, vascular laboratory database was interrogated to identify referred patients with arterial disease who underwent concomitant assessment with ABI and lower limb arterial duplex ultrasound (DUS). Duplex acted as the reference standard. Those who had peripheral computed tomography angiogram (CTA) within 3 months of initial assessment were included in a subgroup analysis to correlate ABI with CTA. The primary end point was the sensitivity and specificity of ABI compared with DUS as the reference standard. Results Concomitant assessment was performed in 438 limbs (250 patients) over a 27-month period. The ABI was normal (0.9 to 1.4) in 196 limbs (44.9%) and abnormal in the remaining 241 limbs (55.1%). False-positive results occurred in 83 out of 241 limbs (34.4%), and false-negative results occurred in 54 limbs out of 196 (27.5%). True-positive results were 158 out of 241 limbs (65.6%), whereas true-negative results were 142 out of 196 limbs (72.4%). ABI using DUS as a benchmark identified a sensitivity for peripheral artery disease of 72.3% and a specificity of 69.3%. Concomitant CTA imaging was available in 200 limbs. The sensitivity and specificity of ABI correlated with CTA were 65.5 and 68.8%, respectively. Conclusion ABIs have a moderate predictive value in the diagnosis of LEAD. Normal range outcomes cannot be taken to infer the absence of LEAD and, as such, further arterial imaging in the form of DUS or angiography should be strongly considered in those with suspected underlying disease requiring intervention. Further noninvasive tests such as exercise studies or pulse volume waveforms should be considered, if diagnostic uncertainty exists, in those requiring nonoperative intervention and risk factor control.

Therapeutic Drug Monitoring: Psychotropic Drugs

1989 ◽  
Vol 2 (6) ◽  
pp. 403-415 ◽  
Author(s):  
Randall D. Seifert
Keyword(s):  
Therapeutic Drug Monitoring ◽  
Drug Monitoring ◽  
Antipsychotic Drugs ◽  
Practical Knowledge ◽  
Therapeutic Monitoring ◽  
Therapeutic Drug ◽  
Clinical Use ◽  
Careful Patient ◽  
Recovery Expectations ◽  
Positive Results

The therapeutic monitoring of patients who take antipsychotic drugs can be both challenging and rewarding. Antipsychotics have been in clinical use for over 30 years; yet, their complex pharmacology is not fully understood and parallels our infant knowledge of human brain chemistry. The art of successful therapeutic drug monitoring depends on the clinician's knowledge of basic pharmacology, an understanding of psychiatric disorders, and a sensitivity for careful patient observation. In addition, a thorough history, well thought out goals, and reasonable recovery expectations are essential. Antipsychotic drugs are never curative and should be used judiciously for indications where positive results outweigh the risks of adverse effects. This article will provide the reader with sound, practical knowledge of how to monitor these drugs in any clinical setting. © 1989 by W.B. Saunders Company.

scholarly journals The Interpretation of Studies Using the Reading Level Design

1989 ◽  
Vol 21 (4) ◽  
pp. 413-424 ◽  
Author(s):  
Usha Goswami ◽  
Peter Bryant
Keyword(s):  
Reading Disability ◽  
Reading Level ◽  
Chronological Age ◽  
Control Group ◽  
Causal Factors ◽  
Negative Results ◽  
Level Design ◽  
Positive Results

Recent research in reading disability has been strengthened by the addition of a reading level (RL) control group to the more traditional chronological age (CA) control group. However, caution is required in interpreting results from these two kinds of control. Only positive results in a RL match and negative results in a CA match are interpretable; negative results in a CA match and positive results in a RL match are not. Furthermore, the RL control group cannot be used to unambiguously determine between specific deficit and developmental lag interpretations of reading disability. It is argued that the use of the RL control can only ever be a first step in research aimed at delineating the causal factors in reading backwardness.

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