scholarly journals Evaluation of “Caterina assay”: An Alternative Tool to the Commercialized Kits Used for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Identification

Pathogens ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 325
Author(s):  
Germano Orrù ◽  
Alessandra Scano ◽  
Sara Fais ◽  
Miriam Loddo ◽  
Mauro Giovanni Carta ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Clinical Microbiology ◽  
Early Stage ◽  
Low Cost ◽  
High Specificity ◽  
N Gene ◽  
Molecular Test ◽  
Specificity And Sensitivity ◽  
Human Coronaviruses ◽  
Commercial Kits

Here we describe the first molecular test developed in the early stage of the pandemic to diagnose the first cases of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in Sardinian patients in February–March 2020, when diagnostic certified methodology had not yet been adopted by clinical microbiology laboratories. The “Caterina assay” is a SYBR®Green real-time reverse-transcription polymerase chain reaction (rRT-PCR), designed to detect the nucleocapsid phosphoprotein (N) gene that exhibits high discriminative variation RNA sequence among bat and human coronaviruses. The molecular method was applied to detect SARS-CoV-2 in nasal swabs collected from 2110 suspected cases. The study article describes the first molecular test developed in the early stage of the declared pandemic to identify the coronavirus disease 2019 (COVID-19) in Sardinian patients in February–March 2020, when a diagnostic certified methodology had not yet been adopted by clinical microbiology laboratories. The assay presented high specificity and sensitivity (with a detection limit ≥50 viral genomes/μL). No false-positives were detected, as confirmed by the comparison with two certified commercial kits. Although other validated molecular methods are currently in use, the Caterina assay still represents a valid and low-cost detection procedure that could be applied in countries with limited economic resources.

scholarly journals Serum Proteomic Fingerprints of Adult Patients with Severe Acute Respiratory Syndrome

2006 ◽  
Vol 52 (3) ◽  
pp. 421-429 ◽  
Author(s):  
Ronald TK Pang ◽  
Terence CW Poon ◽  
KC Allen Chan ◽  
Nelson LS Lee ◽  
Rossa WK Chiu ◽  
...  
Keyword(s):  
Viral Load ◽  
Severe Acute Respiratory Syndrome ◽  
Roc Curve ◽  
Serum Proteins ◽  
Early Stage ◽  
High Specificity ◽  
Roc Curves ◽  
Roc Curve Analysis ◽  
Outbreak Period ◽  
Specificity And Sensitivity

Abstract Background: Severe acute respiratory syndrome (SARS) is an emerging infectious disease caused by a new coronavirus strain, SARS-CoV. Specific proteomic patterns might be present in serum in response to the infection and could be useful for early detection of the disease. Methods: Using surface-enhanced laser desorption/ionization (SELDI) ProteinChip technology, we profiled and compared serum proteins of 39 patients with early-stage SARS infection and 39 non-SARS patients who were suspected cases during the SARS outbreak period. Proteomic patterns associated with SARS were identified by bioinformatic and biostatistical analyses. Features of interest were then purified and identified by tandem mass spectrometry. Results: Twenty proteomic features were significantly different between the 2 groups. Fifteen were increased in the SARS group, and 5 were decreased. Their concentrations were correlated with 2 or more clinical and/or biochemical variables. Two were correlated with the SARS-CoV viral load. Hierarchical clustering analysis showed that a majority of the SARS patients (95%) had similar serum proteomic profiles and identified 2 subgroups with poor prognosis. ROC curve analysis identified individual features as potential biomarkers for SARS diagnosis (areas under ROC curves, 0.733–0.995). ROC curve areas were largest for an N-terminal fragment of complement C3c α chain (m/z 28 119) and an internal fragment of fibrinogen α-E chain (m/z 5908). Immunoglobulin κ light chain (m/z 24 505) positively correlated with viral load. Conclusions: Specific proteomic fingerprints in the sera of adult SARS patients could be used to identify SARS cases early during onset with high specificity and sensitivity.

scholarly journals Cytohistopathological Study of Salivary Gland Lesions in Bundelkhand Region, Uttar Pradesh, India

2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
Anita Omhare ◽  
Sanjeev Kumar Singh ◽  
Jitendra Singh Nigam ◽  
Ankit Sharma
Keyword(s):  
Salivary Gland ◽  
Low Cost ◽  
Uttar Pradesh ◽  
Diagnostic Technique ◽  
High Specificity ◽  
Turnaround Time ◽  
Malignant Tumours ◽  
Female Ratio ◽  
Specificity And Sensitivity ◽  
Benign Tumours

Background. FNAC is a useful method for evaluating suspicious salivary glands lesions due to its low cost, minimum morbidity, rapid turnaround time, high specificity, and sensitivity. Aim. To know the frequency of the salivary gland lesions and cytohistological correlation in the Jhansi region, Uttar Pradesh, India. Material and Methods. In present study 124 cases were included and cytohistological correlation was made in 86 cases only. FNA was performed by using a 23/24-gauge needle without local anaesthesia. Air dried and 95% ethyl alcohol fixed wet smears were stained with Giemsa stain and Papanicolaou stain, respectively. Paraffin embedded tissue sections were stained with Haematoxylin and Eosin. Results. Parotid gland was the most commonly involved salivary gland. The commonest age group was 20 to 29 years, 30 to 39 years, and 60 to 69 years for nonneoplastic lesions, benign tumours, and malignant tumours, respectively. The overall male to female ratio was 1.17 : 1. The diagnostic accuracy of FNAC was 100%, 93.3%, and 88.2% for nonneoplastic lesions, benign tumours, and malignant tumours, respectively. Conclusion. The high accuracy, sensitivity, and specificity of FNAC confirm that preoperative cytology is a useful, quick, reliable diagnostic technique for rapid diagnosis and suitable for developing countries.

scholarly journals The Use of Biomarkers in Early Diagnostics of Pancreatic Cancer

2018 ◽  
Vol 2018 ◽  
pp. 1-10 ◽  
Author(s):  
Lumir Kunovsky ◽  
Pavla Tesarikova ◽  
Zdenek Kala ◽  
Radek Kroupa ◽  
Petr Kysela ◽  
...  
Keyword(s):  
Radical Surgery ◽  
Early Stage ◽  
Locally Advanced ◽  
High Specificity ◽  
Cost Effective ◽  
Ductal Adenocarcinoma ◽  
Specificity And Sensitivity ◽  
Solid Malignancies ◽  
Late Detection ◽  
Resistance To Chemotherapy

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal solid malignancies with increasing incidence. The poor prognosis is due to the aggressive nature of the tumor, late detection, and the resistance to chemotherapy and radiotherapy. A radical surgery procedure is the only treatment that has been shown to improve the 5-year survival rate to 20-25%. However, the majority of patients (80-85%) are diagnosed with locally advanced or metastatic disease and just 15-20% patients are diagnosed in an early stage allowing them to undergo the potentially curative surgical resection. The early detection of PDAC without the use of invasive methods is challenging and discovery of a cost-effective biomarker with high specificity and sensitivity could significantly improve the treatment and survival in these patients. In this review, we summarize current and newly examined biomarkers in early PDAC detection.

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 SARS-CoV-2 S1 and N-Based Serological Assays Reveal Rapid Seroconversion and Induction of Specific Antibody Response in COVID-19 Patients

2020 ◽  
Author(s):  
Abdullah Algaissi ◽  
Mohamed A. Alfaleh ◽  
Sherif 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

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. 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 Recent Progress in Nanomaterials Modified Electrochemical Biosensors for the Detection of MicroRNA

Micromachines ◽  
2021 ◽  
Vol 12 (11) ◽  
pp. 1409
Author(s):  
Sze Shin Low ◽  
Daizong Ji ◽  
Wai Siong Chai ◽  
Jingjing Liu ◽  
Kuan Shiong Khoo ◽  
...  
Keyword(s):  
Electrochemical Biosensor ◽  
Sequence Similarity ◽  
Low Cost ◽  
High Specificity ◽  
Disease Diagnosis ◽  
Electrochemical Biosensors ◽  
High Sequence Similarity ◽  
Aberrant Expression ◽  
Specificity And Sensitivity ◽  
Different Types

MicroRNAs (miRNAs) are important non-coding, single-stranded RNAs possessing crucial regulating roles in human body. Therefore, miRNAs have received extensive attention from various disciplines as the aberrant expression of miRNAs are tightly related to different types of diseases. Furthermore, the exceptional stability of miRNAs has presented them as biomarker with high specificity and sensitivity. However, small size, high sequence similarity, low abundance of miRNAs impose difficulty in their detection. Hence, it is of utmost importance to develop accurate and sensitive method for miRNA biosensing. Electrochemical biosensors have been demonstrated as promising solution for miRNA detection as they are highly sensitive, facile, and low-cost with ease of miniaturization. The incorporation of nanomaterials to electrochemical biosensor offers excellent prospects for converting biological recognition events to electronic signal for the development of biosensing platform with desired sensing properties due to their unique properties. This review introduces the signal amplification strategies employed in miRNA electrochemical biosensor and presents the feasibility of different strategies. The recent advances in nanomaterial-based electrochemical biosensor for the detection of miRNA were also discussed and summarized based on different types of miRNAs, opening new approaches in biological analysis and early disease diagnosis. Lastly, the challenges and future prospects are discussed.

scholarly journals Tips for a reduction of false positives in manual RT-PCR diagnostics of SARS-CoV-2

Bionatura ◽  
2021 ◽  
Vol 3 (3) ◽  
pp. 1948-1954
Author(s):  
Francisco J. Alvarez ◽  
Mariela Perez-Cardenas ◽  
Marco Gudiño ◽  
Markus P. Tellkamp
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
High Specificity ◽  
False Positives ◽  
Rt Pcr ◽  
Manual Operation ◽  
Laboratory Techniques ◽  
Specificity And Sensitivity ◽  
The World ◽  
Pcr Diagnostics ◽  
Positive Results

RT-PCR is the standard gold technique for testing the presence of RNA of the coronavirus causing Severe Acute Respiratory Syndrome (SARS-CoV-2) due to its high specificity and sensitivity. Despite its general use and reliability, no lab in the world is immune to the generation of false positives. These errors cause a loss of confidence in the technique's power and damage the image of laboratories. More importantly, they can take a toll on tested individuals and have economic, psychological, and health-associated effects. Most false positives are caused during a manual operation inside the laboratory. However, not much has been published about the errors associated with particular laboratory techniques used to detect the virus since the beginning of the actual pandemic. This work precisely reflects on events that occur during manual RT-PCR diagnostics in a COVID-19 laboratory, providing tips for reducing false-positive results.

Detection of early-stage pancreatic carcinoma

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. 4613-4613
Author(s):  
D. Gold ◽  
D. E. Modrak ◽  
G. Newsome ◽  
Z. Karanjawala ◽  
R. Hruban ◽  
...  
Keyword(s):  
Pancreatic Carcinoma ◽  
Early Stage ◽  
High Specificity ◽  
Antigen Expression ◽  
Average Concentration ◽  
Precursor Lesions ◽  
Roc Curve Analysis ◽  
Cystic Neoplasms ◽  
Specificity And Sensitivity ◽  
Labeling Pattern

4613 Background: Invasive pancreatic carcinoma is a virtually lethal disease, mostly because of the failure to detect it at a sufficiently early timepoint for successful treatment. Our laboratory has identified a unique biomarker detected by MAb PAM4 that shows high specificity for a mucin glycoprotein expressed by pancreatic carcinoma (PC). While identified in almost 90% of PC and its precursor lesions, the antigen is not detectable in normal pancreas. We are investigating this biomarker for the early detection of PC. Methods: Both immunohistochemical (IHC) and enzyme immunoassay (EIA) were employed for detection and/or quantitation of PAM4-mucin in tissue and sera, respectively. Results: We have extended our prior IHC results with precursor lesions (Clin Cancer Res 2007;13:7380–7); PAM4 gave an intense, diffuse labeling pattern in 81% of mucinous cystic neoplasms (MCN), with an additional 11% showing a focal pattern (n=27). Thus, a total of 92% of MCN showed evidence of PAM4-antigen expression. Of interest, a difference in the labeling pattern was observed in association with the grade of dysplasia, providing easy identification of MCN with high- grade dysplasia. We previously reported use of an EIA for quantitation of PAM4-antigen in sera. The assay demonstrated a sensitivity and specificity of 77% and 95%, respectively, for identification of PC (J Clin Oncol, 2006;24:252–8). We have now confirmed these results in a set of serum specimens (n= 49 PC, 13 normal) for which staging information was available. Overall specificity and sensitivity were 82% and 85%, respectively, calculated by ROC curve analysis (AUC=0.878±0.045; 95% CI=0.769–0.947). Although only a small number of specimens were from patients with stage I disease (n=12), 92% of these were above the cutoff value for positive response. A correlation was observed for average concentration of antigen in the circulation with stage of disease (R2=0.988). Conclusions: IHC and EIA results indicate that PAM4 identifies a biomarker for PC that is present at the earliest stages of neoplastic transformation, thus warranting controlled analyses of larger specimen numbers. (Supported in part by USPHS grant CA096924 from the NIH.) [Table: see text]

scholarly journals Development of NCOVID-19 Colloidal Gold Immunochromatographic Test Strip

2021 ◽  
Vol 10 (1) ◽  
pp. 1
Author(s):  
Wenyi Liu ◽  
Zhaohua Li
Keyword(s):  
Colloidal Gold ◽  
Low Cost ◽  
Virus Detection ◽  
High Specificity ◽  
Accurate Method ◽  
Rapid Screening ◽  
Nucleic Acid Detection ◽  
Specificity And Sensitivity ◽  
Control Serum

<div><p>Purpose: To establish a fast, simple and accurate method and immunoassay test card for the detection of new coronavirus (nCOVID-19) antigen. Methods: In this study, colloidal gold immunochromatography technology was used to detect nCOVID-19 virus antigens through the sandwich method. At the same time, the preparation plan of colloidal gold was improved, and the application of rapid immune-diagnosis technology in other fields was developed. In this study, purified recombinant nCOVID-19 nucleocapsid protein is used as the antigen to prepare murine monoclonal antibodies. The BN02 antibody produced by the mouse is used as the detection antibody to couple with colloidal gold, forming a gold-labeled complex probe. BN9m1 is used as the coating antibody for the C-line, and ProA is used for the T-line. The polymerization of colloidal gold particles enables us to detect the new coronavirus antigen’s appearance. Thus an in vitro rapid detection kit for virus detection can be made. Results: The positive detection rate of the antigen quality control serum with this colloidal gold reagent was 100%. The specificity was 100%, and the sensitivity was 1ng/ml.  Conclusion: The nCOVID-19 antigen detection reagent (colloidal gold method) developed in this research has high specificity and sensitivity, and can be used in conjunction with nucleic acid detection. As a means of detecting nCOVID-19, it can achieve qualitative and rapid screening of samples with advantage such as accuracy, repeatability, and low cost.</p></div>

scholarly journals Potential Markers for Detection and Monitoring of Ovarian Cancer

2011 ◽  
Vol 2011 ◽  
pp. 1-17 ◽  
Author(s):  
Brandon J. D. Rein ◽  
Sajal Gupta ◽  
Rima Dada ◽  
Joelle Safi ◽  
Chad Michener ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Early Detection ◽  
Early Stage ◽  
Survival Rates ◽  
High Specificity ◽  
Protein Markers ◽  
Specificity And Sensitivity ◽  
Novel Biomarkers ◽  
New Biomarkers ◽  
Current Screening

This paper reviews current screening techniques as well as novel biomarkers and their potential role in early detection of ovarian cancer. Ovarian cancer is one of the most common reproductive cancers and has the highest mortality rate amongst gynecologic cancers. Because most ovarian cancer diagnoses occur in the late stages of the disease, five-year survival rates fall below 20%. To improve survival rates and to lower mortality rates for ovarian cancer, improved detection at early stages of the disease is needed. Current screening approaches include tumor markers, ultrasound, or a combination. Efforts are underway to discover new biomarkers of ovarian cancer in order to surmount the obstacles in early-stage diagnosis. Among serum protein markers, HE4 and mesothelin can augment CA125 detection providing higher sensitivity and specificity due to the presence of these proteins in early-stage ovarian cancer. Detection testing that includes methylation of the MCJ gene and increased expression of vascular endothelial growth factor is correlated to poor prognosis and may predict patient survival outcome. Detection testing of biomarkers with long-term stability and combination panels of markers, will likely lead to effective screening strategies with high specificity and sensitivity for early detection of ovarian cancer.

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