scholarly journals Improved Sensitivity of Allergen Detection by Immunoaffinity LC-MS/MS Using Ovalbumin as a Case Study

Foods ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 2932
Author(s):  
Martin Röder ◽  
Claudia Wiacek ◽  
Frauke Lankamp ◽  
Jonathan Kreyer ◽  
Wolfgang Weber ◽  
...  
Keyword(s):  
Food Processing ◽  
False Negative ◽  
Detection Methods ◽  
Food Allergies ◽  
Negative Results ◽  
False Negative Results ◽  
Allergen Detection ◽  
Current Mass ◽  
Egg Powder

Food allergies are caused by severe hypersensitivity to specific food allergens such as the egg protein ovalbumin. It is therefore important to test food products for the presence of allergens to protect allergic people from accidental ingestion. For egg detection, ELISA is the only reasonable commercially available test format, although the recognition of target allergens can be affected by food processing, which may lead to false negative results. Current mass spectrometry-based detection methods may overcome this issue, but these approaches are often less sensitive. Here we combined the advantages of antibody-based and MS-based methods by developing an immunoaffinity LC-MS/MS technique to detect the common egg allergen Gal d 2. We investigated the principal functionality of this method with incurred cookie material containing whole egg powder. We found that the new method matched easily the sensitivity of egg specific ELISA tests. Further western blot experiments indicated that this strategy may be unaffected by food processing, providing an important alternative strategy for the detection and quantification of allergens in food.

scholarly journals Emerging approaches for detection of methylation sites in RNA

Open Biology ◽  
2018 ◽  
Vol 8 (9) ◽  
pp. 180121 ◽  
Author(s):  
Anna Ovcharenko ◽  
Andrea Rentmeister
Keyword(s):  
High Throughput Sequencing ◽  
De Novo ◽  
Direct Detection ◽  
False Negative ◽  
Detection Methods ◽  
Rna Modifications ◽  
Negative Results ◽  
Third Generation Sequencing ◽  
False Negative Results ◽  
Generation Sequencing

RNA methylations play a significant regulatory role in diverse biological processes. Although the transcriptome-wide discovery of unknown RNA methylation sites is essential to elucidate their function, the development of a bigger variety of detection approaches is desirable for multiple reasons. Many established detection methods for RNA modifications heavily rely on the specificity of the respective antibodies. Thus, the development of antibody-independent transcriptome-wide methods is beneficial. Even the antibody-independent high-throughput sequencing-based methods are liable to produce false-positive or false-negative results. The development of an independent method for each modification could help validate the detected modification sites. Apart from the transcriptome-wide methods for methylation detection de novo , methods for monitoring the presence of a single methylation at a determined site are also needed. In contrast to the transcriptome-wide detection methods, the techniques used for monitoring purposes need to be cheap, fast and easy to perform. This review considers modern approaches for site-specific detection of methylated nucleotides in RNA. We also discuss the potential of third-generation sequencing methods for direct detection of RNA methylations.

Variables affecting laboratory diagnosis of acute rickettsial infection

2018 ◽  
Vol 39 (4) ◽  
pp. 220
Author(s):  
Cecilia Kato
Keyword(s):  
Molecular Detection ◽  
False Negative ◽  
Laboratory Diagnosis ◽  
Acute Stage ◽  
Detection Methods ◽  
Rickettsial Infection ◽  
Negative Results ◽  
Enhanced Sensitivity ◽  
False Negative Results ◽  
Antibody Titres

The reference standard for the confirmation of a recent rickettsial infection is by the observation of a four-fold or greater rise in antibody titres when testing paired acute and convalescent (two to four weeks after illness resolution) sera by serological assays (Figure 1). At the acute stage of illness, diagnosis is performed by molecular detection methods most effectively on DNA extracted from tissue biopsies (eschars, skin rash, and organs) or eschar swabs. Less invasive and more convenient samples such as blood and serum may also be used for detection; however, the low number of circulating bacteria raises the possibility of false negative results. Optimal sampling practices and enhanced sensitivity must therefore be considered in order to provide a more accurate laboratory diagnosis.

scholarly journals An appraisal of various pathogen detection methods in eggs and poultry

2020 ◽  
Vol 1 (2) ◽  
pp. 93-97
Author(s):  
A. A. P. Milton ◽  
G. Bhuvana Priya ◽  
K. M. Momin ◽  
M. Angappan ◽  
Samir Das ◽  
...  
Keyword(s):  
Pathogen Detection ◽  
False Negative ◽  
Foodborne Pathogen ◽  
Detection Methods ◽  
Immunological Methods ◽  
Culturable Bacteria ◽  
Product Recalls ◽  
Poultry Products ◽  
Negative Results ◽  
False Negative Results

Abstract: To ensure safety in egg and poultry products, timely detection of pathogenic microbes is of paramount importance. This review offers an appraisal of different routinely used and novel emerging pathogen detection methods in egg, poultry and their products. Timely detection of pathogens is decisive to curtail outbreak risks, reduce hospitalisation, and provide product assurance. It will also reduce the cost of holding food products in cold storage and reduces product recalls. Some crucial issues need to be taken care of in choosing or developing a foodborne pathogen detection method. They are requirement of costly or sophisticated equipment, portability, trained personnel, viable but non-culturable bacteria (may give false-negative results), dead microbes (may give false-positive results), stressed or sub-lethally damaged bacteria and slow-growing microbes (require enrichment). In this review, the focus has been given on culture-based methods, nucleic acid-based methods, immunological methods and biosensor based methods. Keywords: Egg; poultry; detection methods; product assurance; safety.

scholarly journals COVID-19: Test, Test and Test

2020 ◽  
Vol 9 (1) ◽  
pp. 1
Author(s):  
Fatima A Saleh ◽  
Aleen Sleem
Keyword(s):  
Diagnostic Testing ◽  
False Negative ◽  
Detection Methods ◽  
Nucleic Acid Detection ◽  
Rt Pcr ◽  
Negative Results ◽  
Immunological Tests ◽  
False Negative Results ◽  
Different Types ◽  
Polymerase Chain

A new virus was identified in late December 2019 when China reported the first cases of pneumonia in Wuhan, and a global COVID-19 pandemic followed. The world was not late to respond, with a number of sweeping measures ranging from social distancing protocols, stringent hygienic practices, and nation-wide lockdowns, as well as COVID-19 testing campaigns in an attempt to prevent the transmission of the disease and contain the pandemic. Currently, different types of diagnostic testing have been adopted globally, such as nucleic acid detection tests, immunological tests and imaging approaches; however, real-time reverse transcriptase–polymerase chain reaction (RT-PCR) remains the “gold standard” for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Pre-analytical factors, such as specimen selection and collection, are crucial for RT-PCR, and any suboptimal collection may contribute to false-negative results. Herein, we address some of the specimen types that have been used in molecular detection methods for COVID-19. However, the pandemic is still evolving, and information might change as more studies are conducted.

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.

Scoring System for the Diagnosis of COVID-19

2020 ◽  
Vol 13 (1) ◽  
pp. 413-414 ◽  
Author(s):  
Mohamed Farouk Allam
Keyword(s):  
Scoring System ◽  
Clinical Symptoms ◽  
False Negative ◽  
Nasopharyngeal Swab ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Negative Results ◽  
False Negative Results ◽  
Symptoms And Signs

Due to the international spread of COVID-19, the difficulty of collecting nasopharyngeal swab specimen from all suspected patients, the costs of RT-PCR and CT, and the false negative results of RT-PCR assay in 41% of COVID-19 patients, a scoring system is needed to classify the suspected patients in order to determine the need for follow-up, home isolation, quarantine or the conduction of further investigations. A scoring system is proposed as a diagnostic tool for suspected patients. It includes Epidemiological Evidence of Exposure, Clinical Symptoms and Signs, and Investigations (if available). This scoring system is simple, could be calculated in a few minutes, and incorporates the main possible data/findings of any patient.

Detection of Streptococcus pneumoniae in Sputum Samples by Real- Time PCR.

2020 ◽  
Vol 18 ◽  
Author(s):  
Pegah Shakib ◽  
Mohammad Reza Zolfaghari
Keyword(s):  
Streptococcus Pneumoniae ◽  
Real Time ◽  
Real Time Pcr ◽  
False Negative ◽  
Cross Sectional Study ◽  
Laboratory Culture ◽  
Cross Sectional ◽  
Negative Results ◽  
False Negative Results ◽  
Pcr Method

Background: Conventional laboratory culture-based methods for diagnosis of Streptococcus pneumoniae are time-consuming and yield false negative results. Molecular methods including real-time (RT)-PCR rapid methods and conventional PCR due to higher sensitivity and accuracy have been replaced instead traditional culture assay. The aim of the current study was to evaluate lytA gene for detection of Streptococcus pneumoniae in the cerebrospinal fluid of human patients with meningitis using real-time PCR assay. Material and Methods: In this cross-sectional study, a total of 30 clinical specimens were collected from patients in a period from September to December 2018. In order to evaluate the presence of lytA gene, conventional and real-time PCR methods were used without culture. Results: From 30 sputum samples five (16.66%) isolates were identified as S. pneumoniae by lytA PCR and sequencing. Discussion: In this research, an accurate and rapid real-time PCR method was used, which is based on lytA gene for diagnosis of bacteria so that it can be diagnosed. Based on the sequencing results, the sensitivity for detection of lytA gene was 100% (5/5).

COVID-19: how nuclear medicine can provide a differential diagnosis in very dubious case

Coronaviruses ◽  
2020 ◽  
Vol 01 ◽  
Author(s):  
Maria Silvia De Feo ◽  
Viviana Frantellizzi ◽  
Giuseppe De Vincentis
Keyword(s):  
Infectious Disease ◽  
Differential Diagnosis ◽  
Ct Scan ◽  
Imaging Technique ◽  
False Negative ◽  
Clinical Signs ◽  
Negative Results ◽  
Infectious Disease Department ◽  
False Negative Results ◽  
99Mtc Hmpao

Background: We present the case of a 55-year-old woman, admitted to the Infectious Disease Department of Policlinico Umberto I, Rome, in mid-March 2020, with suspicion of COVID-19 infection. Objective: The rRT-PCR was negative and the following CT scan, performed to exclude false-negative results and help diagnosis, was inconclusive. Methods: It was decided to submit the patient to 99mTc-HMPAO-labelled leukocyte scan. Results: This exam led to the diagnosis of infective endocarditis. Conclusion: In the present pandemic scenario, 99mTc-HMPAO-labelled leukocyte scan represents a reliable imaging technique for differential diagnosis with COVID-19 in patients with confusing clinical signs, possible false-negative rRT-PCR results and inconclusive CT scan.

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