Development of test kit for detection of specific IgM to SARS-CoV-2 by immune blotting in the «Line blot» format

2021 ◽  
Vol 66 (8) ◽  
pp. 472-479
Author(s):  
S. G. Mardanly ◽  
A. S. Avdonina
Keyword(s):  
Blood Serum ◽  
Sensitivity And Specificity ◽  
High Sensitivity ◽  
Confirmatory Test ◽  
Clinical Samples ◽  
Etiological Diagnosis ◽  
Healthy Donors ◽  
Test Kit ◽  
Preliminary Study

Test kit for detection of specific IgM to SARS-CoV-2 by immune blotting in the «Line blot» format has been developed. A preliminary study of diagnostic effectivity on clinical samples of blood serum from patients with COVID-19 and healthy donors showed its high sensitivity and specificity. The new test kit allows to detect IgM to all four structural antigens of SARS-CoV-2 and can be used as a confirmatory test to verify indeterminant screening results in laboratory etiological diagnosis of COVID-19.

A Preliminary Evaluation of a Prototype Western Blot Confirmatory Test Kit for Syphilis

1994 ◽  
Vol 5 (6) ◽  
pp. 409-414 ◽  
Author(s):  
H Young ◽  
P J Walker ◽  
D Merry ◽  
A Mifsud
Keyword(s):  
Western Blot ◽  
False Positive ◽  
High Sensitivity ◽  
High Specificity ◽  
Confirmatory Test ◽  
Preliminary Evaluation ◽  
Serological Tests ◽  
Western Blot Test ◽  
Specificity And Sensitivity ◽  
Test Kit

A prototype Western blot kit was evaluated as a confirmatory test for syphilis using 131 sera characterized by other serological tests for syphilis. There were 114 treponemal sera (including 94 cases of early syphilis, 83 of which were untreated) and 17 non-treponemal problem sera (11 gave false positive reactions on screening with the TmpA recombinant antigen enzyme immunoassay (EIA), 3 gave false positive fluorescent treponemal antibody absorbed (FTA-abs) tests, and 3 false positive Captia Syphilis G EIA results). Based on the manufacturer's criteria of reactivity in multiple bands for designating a positive result the Western blot test gave a sensitivity of 99.1% (113/114) and a specificity of 88.2% (15/17) when indeterminate reactions were scored positive and 98.2% (112/114) and 100% (17/17) when indeterminate reactions were scored negative. Sensitivity was high in both treated and untreated infection. Corresponding sensitivities for the TPHA and FTA-abs when equivocal reactions were scored negative were 97.5% (111/114) and 99.1% (113/114). The high sensitivity of the FTA-abs in this study is probably due to the large number of untreated primary infections. Our results with the Western blot, confirm earlier studies using ‘in-house’ test systems and, support a role for a commercial Western blot test in the confirmatory diagnosis of syphilis. Further studies are required to confirm the high specificity and sensitivity of the kit in a larger series including a wider variety of non-treponemal cases as well as patients with untreated and treated infection.

scholarly journals Epigenetic Markers for Molecular Detection of Prostate Cancer

2007 ◽  
Vol 23 (1-2) ◽  
pp. 31-41 ◽  
Author(s):  
Vera L. Costa ◽  
Rui Henrique ◽  
Carmen Jerónimo
Keyword(s):  
Prostate Cancer ◽  
Sensitivity And Specificity ◽  
Prostate Cancer Risk ◽  
High Sensitivity ◽  
Promoter Hypermethylation ◽  
Clinical Samples ◽  
Screening Methods ◽  
Tissue Samples ◽  
Age Related ◽  
Small Set

Prostate cancer is a highly prevalent malignancy, which is clinically silent but curable while organ-confined. Because available screening methods show poor sensitivity and specificity, the development of new molecular markers is warranted. Epigenetic alterations, mainly promoter hypermethylation of cancer-related genes, are common events in prostate cancer and might be used as cancer biomarkers. Moreover, the development of quantitative, high-throughput techniques to assess promoter methylation enabled the simultaneous screening of multiple clinical samples. From the numerous cancer-related genes hypermethylated in prostate cancer only a few proved to be strong candidates to become routine biomarkers. This small set of genes includesGSTP1,APC,RARβ2,Cyclin D2,MDR1, andPTGS2. Single and/or multigene analyses demonstrated the feasibility of detecting early prostate cancer, with high sensitivity and specificity, in body fluids (serum, plasma, urine, and ejaculates) and tissue samples. In addition, quantitative hypermethylation of several genes has been associated with clinicopathologic features of tumor aggressiveness, and also reported as independent prognostic factor for relapse. The identification of age-related methylation at specific loci and the differential frequency of methylation among ethnical groups, also provided interesting data linking methylation and prostate cancer risk. Although large trials are needed to validate these findings, the clinical use of these markers might be envisaged for the near future.

scholarly journals Real-time RT-PCR diagnostics of virus causing COVID-19

2020 ◽  
Vol 13 (1) ◽  
pp. 52-63 ◽  
Author(s):  
E. V. Goncharova ◽  
A. E. Donnikov ◽  
V. V. Kadochnikova ◽  
S. A. Morozova ◽  
M. N. Boldyreva ◽  
...  
Keyword(s):  
Real Time ◽  
Sensitivity And Specificity ◽  
High Sensitivity ◽  
Medical Product ◽  
World Health ◽  
Differential Diagnostics ◽  
Clinical Samples ◽  
Rt Pcr ◽  
Study Results ◽  
Pcr Diagnostics

Aim: the study was aimed to develop a reagent kit for the real-time RT-PCR diagnostics of virus causing COVID-19.Materials and Methods. Three target sites were chosen in the genome SARS-CoV-2. The testing included 220 samples, 48 artificially created positive samples (made from patients’ biomaterial) and 172 clinical samples (scrapes from nasal and pharyngeal cavities, bronchoalveolar lavage, expectoration, endotracheal/nasopharyngeal aspirate, feces, post-mortem material), obtained from two medical centers. Preliminary, the obtained biomaterial was analyzed with a reagent kit of comparison. The evaluation was performed with a confidential interval CI 95%. The calculation of CI for the sensitivity and specificity was made based on the distribution of χ2.Results. The authors developed a technology of novel coronavirus infection (COVID-19) real-time RT-PCR diagnostics for the application in practical healthcare and proposed the variants of testing at all the stages (preanalytical, analytical, and post-analytical, including automated results processing). The proposed reagent kit meets the requirements of the World Health Organization and the Ministry of Healthcare of the Russian Federation. The study results demonstrated high sensitivity and specificity. The sensitivity was 100% (95% CI) 95.6–100%; the specificity was 100% (95% CI) 96.7–100%.Conclusion. The proposed reagent kit was registered in the RF as a medical product; the registration certificate No. RZN 2020/9948 dated 01.04.2020. The application of the reagent kit in network laboratories will provide patients with access to testing for the virus causing COVID-19 and contribute to quick differential diagnostics, improvement of pandemic control, and accurate statistics on the spread of the virus. 

Whole-genome sequence-guided PCR for the rapid identification of thePseudomonas aeruginosa ST175 high-risk clone directly from clinical samples

2020 ◽  
Author(s):  
Gabriel Cabot ◽  
Paula Lara-Esbrí ◽  
Xavier Mulet ◽  
Antonio Oliver
Keyword(s):  
High Risk ◽  
Sensitivity And Specificity ◽  
Genome Sequence ◽  
Treatment Strategies ◽  
High Sensitivity ◽  
Whole Genome Sequence ◽  
Clinical Samples ◽  
Whole Genome ◽  
Pcr Assay ◽  
Pcr Targeting

Abstract Objectives Pseudomonas aeruginosa frequently show MDR/XDR profiles, which are associated with worldwide-disseminated high-risk clones (HRCs). We developed a PCR assay for the detection in clinical samples of ST175, an HRC that is widespread in European countries. Methods The whole-genome sequence was obtained for one ST175 isolate using a PacBio RSII sequencer. Reads from multiple isolates belonging to ST175 and the PAO1 reference strain were mapped against the ST175 genome to identify potentially specific regions. Once curated, using the BLAST database to search for the presence of those regions in any other organism, we designed a specific PCR for the detection of ST175. Results Assembly of the ST175 PacBio-sequenced genome resulted in three contigs with a total length of 7 087 985 bases, encoding 6566 coding sequences. Specific regions for ST175 genomes were detected and a PCR targeting a 318 bp fragment located within a 3177 bp ORF coding for a putative reverse transcriptase was designed. The PCR test was first evaluatedin silico against 229 XDRP. aeruginosa genomes (73 ST175) from two multicentre studies, yielding 100% sensitivity and specificity. Then, the PCR was evaluatedin vitro in 25 isolates (12 ST175) and in 120 clinical samples (30 urine samples, 30 blood cultures, 30 sputum samples and 30 rectal swabs) of which 10% contained ST175, yielding again 100% sensitivity and specificity. Conclusions The PCR assay developed, showing high sensitivity and specificity for the detection of the ST175 HRC directly from clinical samples, could become a useful tool for guiding infection control and treatment strategies in areas with a high prevalence of this clone.

scholarly journals Evaluation of a Rapid Kit for Detection of IgM against Leptospira in Human

2019 ◽  
Vol 2019 ◽  
pp. 1-4 ◽  
Author(s):  
Mohan Rao ◽  
Fairuz Amran ◽  
Nadia Aqilla
Keyword(s):  
Sensitivity And Specificity ◽  
Predictive Value ◽  
Clinical Manifestations ◽  
Rapid Test ◽  
Febrile Illness ◽  
High Sensitivity ◽  
Confirmatory Test ◽  
Standard Laboratory ◽  
Test Efficiency ◽  
Sensitivity Specificity

Introduction. Leptospirosis is an acute febrile illness, known for its protean clinical manifestations and the challenge in differentiating from other infectious diseases. Standardized confirmatory test is antibody dependent and not accessible by the suburban community. This study measures efficiency of an immune-chromatographic assay, Leptocheck WB, in detecting acute leptospirosis. Methods. A total of 142 sera were used for kit evaluation. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated by comparing rapid kit results with gold standard laboratory, microscopic agglutination test (MAT). Results. We found this rapid kit to have a sensitivity and specificity of 66.6% and 78.9%, respectively, whereas the PPV and NPV of the kit appeared to be 73.3% and 73.2%, respectively. Discussion. Test efficiency of this rapid kit is reasonable. It is specific in detecting leptospiral antibody and assures clinician of accurate diagnosis by having higher PPV and NPV. It is prompt and efficient in comparison with conventional methods in assisting differential diagnosis. High sensitivity and specificity leptospirosis rapid test is indeed a crucial measure to assist the diagnosis of acute undifferentiated febrile illnesses.

scholarly journals Rapid and Visible RPA-Cas12a fluorescence Assay for Accurate Detection of Zoonotic Dermatophytes

2021 ◽  
Author(s):  
Liyang Wang ◽  
Jinyu Fu ◽  
Guang Cai ◽  
Di Zhang ◽  
Shuobo Shi ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Limit Of Detection ◽  
Diagnostic Testing ◽  
High Sensitivity ◽  
Trichophyton Mentagrophytes ◽  
Fluorescence Assay ◽  
Diagnostic Methods ◽  
Clinical Samples ◽  
Microsporum Canis ◽  
Fungal Culture

Background: Dermatophytosis is an infectious disease of global significance caused by several fungal species, which affects the hair, nails, or superficial layers of the skin. The most common zoonotic dermatophytes are Microsporum canis, Nannizzia gypsea and Trichophyton mentagrophytes. Wood's lamp examination, microscopic identification and fungal culture are the main conventional diagnostic methods used in clinics. Less common methods are dermatophyte PCR and biopsy/histopathology. However, these methods also have limitations for providing both accuracy and timely on-site detection. The recent development of CRISPR-based diagnostic platform provides the possibility of a rapid, accurate, and portable diagnostic tool, which has huge potential for clinical applications. Objectives: The purpose of this study is to establish a molecular method for rapid and accurate diagnosis of clinical dermatophytes, which can accelerate clinical diagnostic testing and help timely treatment. Methods: In this paper, we design a Cas12a-based assay combined with recombinase polymerase amplification (RPA) to differentiate three main zoonotic dermatophytes. The limit of detection (LOD) is determined by using standard strains. A total of 25 clinical samples (hair and scurf) are identified to evaluate the sensitivity and specificity of this assay. Results: The RPA-Cas12a method showed high sensitivity and specificity (100% and 100%, respectively). The results could be observed directly by naked-eyes, and all tested samples were consistent with fungal culture and sequencing results. Conclusions: Compared with other methods, the RPA-Cas12a-fluorescence assay requires less time (30 minutes) and less complicated equipment, and visible changes can be clearly observed, which is suitable for on-site clinical diagnosis.

scholarly journals Fast diagnostic test for familial Mediterranean fever based on a kinase inhibitor

2020 ◽  
Vol 80 (1) ◽  
pp. 128-132
Author(s):  
Flora Magnotti ◽  
Tiphaine Malsot ◽  
Sophie Georgin-lavialle ◽  
Fatima Abbas ◽  
Amandine Martin ◽  
...  
Keyword(s):  
Familial Mediterranean Fever ◽  
Sensitivity And Specificity ◽  
Kinase Inhibitor ◽  
Gene Dosage ◽  
Characteristic Curve ◽  
High Sensitivity ◽  
Inflammasome Activation ◽  
Clinical Criteria ◽  
Healthy Donors ◽  
Mediterranean Fever

Background and objectiveFamilial Mediterranean fever (FMF) is the most frequent hereditary autoinflammatory disease. Its diagnosis relies on a set of clinical criteria and a genetic confirmation on identification of biallelic pathogenic MEFV variants. MEFV encodes pyrin, an inflammasome sensor. Using a kinase inhibitor, UCN-01, we recently identified that dephosphorylation of FMF-associated pyrin mutants leads to inflammasome activation. The aim of this study was to assess whether quantifying UCN-01-mediated inflammasome activation could discriminate FMF patients from healthy donors (HD) and from patients with other inflammatory disorders (OID).MethodsReal-time pyroptosis and IL-1β secretion were monitored in response to UCN-01 in monocytes from FMF patients (n=67), HD (n=71) and OID patients (n=40). Sensitivity and specificity of the resulting diagnostic tests were determined by receiver operating characteristic curve analyses.ResultsInflammasome monitoring in response to UCN-01 discriminates FMF patients from other individuals. Pyroptosis assessment leads to a fast FMF diagnosis while combining pyroptosis and IL-1β dosage renders UCN-01-based assays highly sensitive and specific. UCN-01-triggered monocytes responses were influenced by MEFV gene dosage and MEFV mutations in a similar way as clinical phenotypes are.ConclusionsUCN-01-based inflammasome assays could be used to rapidly diagnose FMF, with high sensitivity and specificity.

Detector-C: A Blood-based IVD with High Sensitivity and Specificity for Early Detection and Screening of Colorectal Cancer

2010 ◽  
Vol 48 (08) ◽  
Author(s):  
A Rosenthal ◽  
H Köppen ◽  
R Musikowski ◽  
R Schwanitz ◽  
J Behrendt ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Early Detection ◽  
Sensitivity And Specificity ◽  
High Sensitivity

Trends in Diagnosis for Active Tuberculosis Using Nanomaterials

2019 ◽  
Vol 26 (11) ◽  
pp. 1946-1959 ◽  
Author(s):  
Le Minh Tu Phan ◽  
Lemma Teshome Tufa ◽  
Hwa-Jung Kim ◽  
Jaebeom Lee ◽  
Tae Jung Park
Keyword(s):  
Sensitivity And Specificity ◽  
High Efficiency ◽  
Point Of Care ◽  
High Sensitivity ◽  
Signs And Symptoms ◽  
Diagnostic Methods ◽  
Advantages And Disadvantages ◽  
Treatment And Prevention ◽  
Clinical Tools ◽  
Diagnostic Applications

Background:Tuberculosis (TB), one of the leading causes of death worldwide, is difficult to diagnose based only on signs and symptoms. Methods for TB detection are continuously being researched to design novel effective clinical tools for the diagnosis of TB.Objective:This article reviews the methods to diagnose TB at the latent and active stages and to recognize prospective TB diagnostic methods based on nanomaterials.Methods:The current methods for TB diagnosis were reviewed by evaluating their advantages and disadvantages. Furthermore, the trends in TB detection using nanomaterials were discussed regarding their performance capacity for clinical diagnostic applications.Results:Current methods such as microscopy, culture, and tuberculin skin test are still being employed to diagnose TB, however, a highly sensitive point of care tool without false results is still needed. The utilization of nanomaterials to detect the specific TB biomarkers with high sensitivity and specificity can provide a possible strategy to rapidly diagnose TB. Although it is challenging for nanodiagnostic platforms to be assessed in clinical trials, active TB diagnosis using nanomaterials is highly expected to achieve clinical significance for regular application. In addition, aspects and future directions in developing the high-efficiency tools to diagnose active TB using advanced nanomaterials are expounded.Conclusion:This review suggests that nanomaterials have high potential as rapid, costeffective tools to enhance the diagnostic sensitivity and specificity for the accurate diagnosis, treatment, and prevention of TB. Hence, portable nanobiosensors can be alternative effective tests to be exploited globally after clinical trial execution.

Creatinuria and Dynamics of Calcium Metabolism in Children in the Phase of Exacerbation of Bronchial Asthma

2020 ◽  
Vol 16 (1) ◽  
pp. 28-33
Author(s):  
Vitalii B. Kaliberdenko ◽  
Shanmugaraj Kulanthaivel ◽  
Michael V. Shterenshis ◽  
Olga Y. Poleshchuk ◽  
Kadri Mametov ◽  
...  
Keyword(s):  
Creatine Kinase ◽  
Blood Serum ◽  
Bronchial Asthma ◽  
Picric Acid ◽  
Colorimetric Method ◽  
Cellular Level ◽  
Creatine Kinase Activity ◽  
Biological Medium ◽  
Creatinine Concentration ◽  
Test Kit

: Bronchial asthma is one of the most common and severe diseases among children. The phenomenon of creatinuria (CU) in patients with bronchial asthma (BA) has been acknowledged for a relatively long time. Aims: The Aim of the research is to study the level of creatinuria, creatinemia, creatine kinase activity, and the concentration of calcium in biological medium (blood, saliva, urine) in children suffering from an intermittent and persistent form of asthma during the period of exacerbation. Material and methods:: The research consists of 102 children with asthma who were treated in inpatient department in Simferopol Clinic. The intermittent course of asthma was recorded in 49 children and a persistent course of asthma was recorded in 53 children. The subject of study was blood serum and daily urine of observed patients. The level of calcium in the biological medium was studied using the "Filisit" test kit (Dnipro) and the activity of the creatine kinase by test set "Lahma". The levels of creatine and creatinine were determined using a colorimetric method based on a color reaction with picric acid. Results and conclusion: : The analysis testifies that creatinuria in children with persistent BA is caused by the disorder of the phosphorylation process rather than the disorder of creatinin rephosphorylation synthesis, that is testified by the normal creatinine level. In children with persistent BA, there is а decrease of creatinine concentration in the blood serum and urine during the exacerbation period and early post exacerbation period. The low activity of creatinine kinase at the background of creatinine elimination is typical for the children in the phase of exacerbation of persistent form of BA, though its level remains to be sufficient for the synthesis of the necessary amount of creatinine phosphate. Conclusion: The processes of creatinuria and calciuria in children suffering from a persistent form of BA are interdependent, that is testified by the data of correlative analysis. In connection with this, it is possible to consider the change of calcium homeostasis in the pathogenesis of the disease as one of the causes of distributing the creatinine metabolism on the cellular level.

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