Morphologic Identification of Viral Gastroenteritis Caused by a Human Calicivirus Strain Undetected Initially by Molecular or Immunologic Methods

1997 ◽  
Vol 3 (S2) ◽  
pp. 79-80 ◽  
Author(s):  
C. D. Humphrey ◽  
J. S. Noel ◽  
B. L. Liu ◽  
E. M. Rodriguez ◽  
P. R. Lambden ◽  
...  
Keyword(s):  
High Specificity ◽  
Viral Gastroenteritis ◽  
Norwalk Virus ◽  
Igg Antibody ◽  
Virus Identification ◽  
Specificity And Sensitivity ◽  
Stool Specimens ◽  
Polymerase Chain ◽  
Identification And Characterization ◽  
Virus Strains

Molecular and immunologie assays for virus identification and characterization are well known for their high specificity and sensitivity. High specificity occasionally hampers efforts to detect virus strains distantly related to previously recognized types and may allow infectious agents to be undetected when these techniques are used for identification. We describe in this report such an example with an outbreak of foodborne gastroenteritis.Forty-six cases of gastrointestinal illness were reported in May 1994 among adult staff of a Parkville, Md., school after a catered luncheon. Symptoms included stomach cramps, nausea, diarrhea, and vomiting. Seven stool specimens, negative by routine bacteriological screening, and 14 acute-and convalescent-phase sera from patients involved in the outbreak were tested for viruses. The stool specimens were examined by electron microscopy (EM) and reverse transcription-polymerase chain reaction (RT-PCR) designed to detect small round structured viruses (SRSVs). In addition, serum specimens were tested for IgG antibody to recombinant capsid proteins from Norwalk virus (rNV) and Toronto virus (rTV) using a direct enzyme immunoassay (EIA).

Development of Dual Fluorescent Microsphere Immunological Assay for Detection of Pseudorabies Virus gE and gB IgG Antibodies

2020 ◽  
Author(s):  
chihai ji ◽  
Jingyu Wang ◽  
Yuchen Zeng ◽  
Haoming Pan ◽  
Yingfang Wei ◽  
...  
Keyword(s):  
Pseudorabies Virus ◽  
High Specificity ◽  
Antibody Detection ◽  
Economic Losses ◽  
Igg Antibodies ◽  
Wild Type ◽  
Swine Industry ◽  
Igg Antibody ◽  
Fluorescent Microsphere ◽  
Specificity And Sensitivity

Abstract Background Pseudorabies, also known as Aujezsky’s disease, is an acute viral infection caused by pseudorabies virus (PRV). Swine are one of the natural hosts of pseudorabies, therefore, the disease brings huge economic losses to the swine industry. Establishment of a differential diagnosis technique that can distinguish between wild-type infected and vaccinated pigs, and monitor vaccine-induced IgG is crucial for eventual eradication of pseudorabies.Results The aim of this study was to develop a rapid dual detection method for PRV gE and gB protein IgG antibodies with high specificity and sensitivity. PRV gE codons at amino acid residues (aa) 52–238 and gB codons at aa 539–741 were expressed to obtain recombinant PRV gE and gB proteins by pMAL-c5x vector. After purification with Qiagen Ni–NTA agarose affinity, the two proteins were analyzed by SDS-PAGE and immunoblotting assay. Two single fluorescent-microsphere immunoassays (FMIA) were established by coupling two recombinant proteins (gE and gB) with two magnetic microbeads and an effective dual FMIA was developed by integrating the two single assays. Optimal serum dilution for each assay, correlation with other common swine virus-positive sera and comparison with ELISA for two PRV antigens were tested for validation. Compared with ELISA, the specificity and sensitivity were 99.26% and 92.3% for gE IgG antibody detection and 95.74% and 96.3% for gB IgG antibody detection by dual-FMIA.Conclusion We provide a new method for monitoring PRV protective antibody in vaccinated pigs and differentiating wild-type-PRV-infected from vaccinated pigs

scholarly journals An outbreak of viral gastroenteritis associated with consumption of sandwiches: implications for the control of transmission by food handlers

1998 ◽  
Vol 121 (3) ◽  
pp. 615-621 ◽  
Author(s):  
U. D. PARASHAR ◽  
L. DOW ◽  
R. L. FANKHAUSER ◽  
C. D. HUMPHREY ◽  
J. MILLER ◽  
...  
Keyword(s):  
Food Handler ◽  
Common Source ◽  
Viral Gastroenteritis ◽  
Food Handlers ◽  
Rt Pcr ◽  
Source Of Infection ◽  
Food Borne ◽  
Pcr Product ◽  
Stool Specimens ◽  
Polymerase Chain

Although food handlers are often implicated as the source of infection in outbreaks of food-borne viral gastroenteritis, little is known about the timing of infectivity in relation to illness. We investigated a gastroenteritis outbreak among employees of a manufacturing company and found an association (RR=14·1, 95% CI=2·0–97·3) between disease and eating sandwiches prepared by 6 food handlers, 1 of whom reported gastroenteritis which had subsided 4 days earlier. Norwalk-like viruses were detected by electron microscopy or reverse transcriptase-polymerase chain reaction (RT-PCR) in stool specimens from several company employees, the sick food handler whose specimen was obtained 10 days after resolution of illness, and an asymptomatic food handler. All RT-PCR product sequences were identical, suggesting a common source of infection. These data support observations from recent volunteer studies that current recommendations to exclude food handlers from work for 48–72 h after recovery from illness may not always prevent transmission of Norwalk-like viruses because virus can be shed up to 10 days after illness or while exhibiting no symptoms.

scholarly journals Human viral gastroenteritis.

1989 ◽  
Vol 2 (1) ◽  
pp. 51-89 ◽  
Author(s):  
M L Christensen
Keyword(s):  
Electron Microscopic Examination ◽  
Electrolyte Imbalance ◽  
Viral Gastroenteritis ◽  
Norwalk Virus ◽  
Older Children ◽  
Electron Microscopic ◽  
Detection Systems ◽  
Rotavirus Vaccines ◽  
Adolescents And Adults ◽  
Stool Specimens

During the last 15 years, several different groups of fastidious viruses that are responsible for a large proportion of acute viral gastroenteritis cases have been discovered by the electron microscopic examination of stool specimens. This disease is one of the most prevalent and serious clinical syndromes seen around the world, especially in children. Rotaviruses, in the family Reoviridae, and fastidious fecal adenoviruses account for much of the viral gastroenteritis in infants and young children, whereas the small caliciviruses and unclassified astroviruses, and possibly enteric coronaviruses, are responsible for significantly fewer cases overall. In addition to electron microscopy, enzyme immunoassays and other rapid antigen detection systems have been developed to detect rotaviruses and fastidious fecal adenoviruses in the stool specimens of both nonhospitalized patients and those hospitalized for dehydration and electrolyte imbalance. Experimental rotavirus vaccines have also been developed, due to the prevalence and seriousness of rotavirus infection. The small, unclassified Norwalk virus and morphologically similar viruses are responsible for large and small outbreaks of acute gastroenteritis in older children, adolescents, and adults. Hospitalization of older patients infected with these viruses is usually not required, and their laboratory diagnoses have been limited primarily to research laboratories.

scholarly journals Production and Characterization of a Polyclonal Antibody of Anti-rLipL21-IgG againstLeptospirafor Early Detection of Acute Leptospirosis

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Arivudainambi Seenichamy ◽  
Abdul Rani Bahaman ◽  
Abdul Rahim Mutalib ◽  
Siti Khairani-Bejo
Keyword(s):  
Polyclonal Antibody ◽  
Diagnostic Marker ◽  
Bacterial Species ◽  
High Specificity ◽  
Cross Reactivity ◽  
Zoonotic Diseases ◽  
Igg Antibody ◽  
E Coli ◽  
Specificity And Sensitivity

Leptospirosis is one of the zoonotic diseases in animals and humans throughout the world. LipL21 is one of the important surface-exposed lipoproteins in leptospires and the most effective cross protective immunogenic antigen. It is widely considered as a diagnostic marker for leptospirosis. In this study, we evaluated the serodiagnostic potential of LipL21 protein ofLeptospira interrogansserovar Pomona. We have successfully amplified, cloned, and expressed LipL21 inE. coliand evaluated its specificity by immunoblotting. Purified recombinant LipL21 (rLipL21) was inoculated into rabbits for the production of polyclonal antibody. Characterization of the purified IgG antibody against rLipL21 was performed by cross reactivity assay. Only sera from leptospirosis patients and rabbit hyperimmune sera recognized rLipL21 while the nonleptospirosis control sera showed no reaction in immunoblotting. We confirmed that anti-rLipL21-IgG antibody cross reacted with and detected only pathogenic leptospiral species and it did not react with nonpathogenic leptospires and other bacterial species. Results observed showed that anti-rLipL21-IgG antibody has high specificity and sensitivity to leptospires. The findings indicated that the antibody could be used in a diagnostic assay for detection of leptospires or their proteins in the early phase of infection.

scholarly journals Development of a Dual Fluorescent Microsphere Immunological Assay for Detection of Pseudorabies Virus gE and gB IgG Antibodies

Viruses ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 912
Author(s):  
Chihai Ji ◽  
Yingfang Wei ◽  
Jingyu Wang ◽  
Yuchen Zeng ◽  
Haoming Pan ◽  
...  
Keyword(s):  
Pseudorabies Virus ◽  
High Specificity ◽  
Nitrilotriacetic Acid ◽  
Antibody Detection ◽  
Economic Losses ◽  
Igg Antibodies ◽  
Wild Type ◽  
Igg Antibody ◽  
Fluorescent Microsphere ◽  
Specificity And Sensitivity

Pseudorabies, also known as Aujezsky’s disease, is an acute viral infection caused by pseudorabies virus (PRV). Swine are one of the natural hosts of pseudorabies and the disease causes huge economic losses in the pig industry. The establishment of a differential diagnosis technique that can distinguish between wild-type infection and vaccinated responses and monitor vaccine-induced immunoglobulin G(IgG) is crucial for the eventual eradication of pseudorabies. The aim of this study was to develop a rapid dual detection method for PRV gE and gB protein IgG antibodies with high specificity and sensitivity. PRV gE codons at amino acid residues (aa) 52–238 and gB codons at aa 539–741 were expressed to obtain recombinant PRV gE and gB proteins via a pMAL-c5x vector. After purification with Qiagen Ni–nitrilotriacetic acid (NTA) agarose affinity chromatography, the two proteins were analyzed via SDS-PAGE and immunoblotting assays. Two single fluorescent-microsphere immunoassays (FMIAs) were established by coupling two recombinant proteins (gE and gB) to magnetic microbeads, and an effective dual FMIA was developed by integrating the two single assays. Optimal serum dilution for each assay, correlation with other common swine virus-positive sera, and comparison with ELISA for two PRV antigens were tested for validation. Compared with ELISA, the specificity and sensitivity were 99.26% and 92.3% for gE IgG antibody detection, and 95.74% and 96.3% for the gB IgG antibody detection via dual FMIA. We provide a new method for monitoring PRV protective antibodies in vaccinated pigs and differentiating wild-type PRV infection from vaccinated responses simultaneously.

scholarly journals ProbeSpec: batch specificity testing and visualization of oligonucleotide probe sets implemented in ARB

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 1901
Author(s):  
Tim Kahlke ◽  
Paavo Jumppanen ◽  
Ralf Westram ◽  
Guy C.G. Abell ◽  
Levente Bodrossy
Keyword(s):  
Sensitivity And Specificity ◽  
Quantitative Polymerase Chain Reaction ◽  
High Specificity ◽  
Cost Effective ◽  
Marker Genes ◽  
Oligonucleotide Probes ◽  
Cross Hybridization ◽  
Specificity And Sensitivity ◽  
Novel Taxa ◽  
Polymerase Chain

High-throughput molecular methods such as quantitative polymerase chain reaction (qPCR) and environmental microarrays are cost-effective methods for semi-quantitative assessment of bacterial community structure and the identification of specific target organisms. Both techniques rely on short nucleotide sequences, so-called oligonucleotide probes, which require high specificity to the organisms in question to avoid cross-hybridization with non-target taxa. However, designing oligonucleotide probes for novel taxa or marker genes that show sufficient phylogenetic sensitivity and specificity is often time- and labor-intensive, as each probe has to be in-silico tested for its specificity and sensitivity. Here we present ProbeSpec, to our knowledge the first batch sensitivity and specificity estimation and visualization tool for oligonucleotide probes integrated into the widely used ARB software. Using ProbeSpec’s interactive “mismatch threshold” and “clade marked threshold” we were able to reduce the development time of highly specific probes for a recently published environmental oligonucleotide microarray from several months to one week.

scholarly journals Detection of Norwalk virus in stool specimens by reverse transcriptase-polymerase chain reaction and nonradioactive oligoprobes.

1992 ◽  
Vol 30 (12) ◽  
pp. 3151-3157 ◽  
Author(s):  
R De Leon ◽  
S M Matsui ◽  
R S Baric ◽  
J E Herrmann ◽  
N R Blacklow ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Norwalk Virus ◽  
Chain Reaction ◽  
Stool Specimens ◽  
Polymerase Chain

scholarly journals Application of PCR-ELISA in Molecular Diagnosis

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Mei Jean Sue ◽  
Swee Keong Yeap ◽  
Abdul Rahman Omar ◽  
Sheau Wei Tan
Keyword(s):  
Detection Limit ◽  
High Specificity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specificity And Sensitivity ◽  
Pcr Product ◽  
Diagnostic Time ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Agricultural Industries ◽  
Analytical Time

Polymerase chain reaction-enzyme linked immunosorbent assay (PCR-ELISA) is an immunodetection method that can quantify PCR product directly after immobilization of biotinylated DNA on a microplate. This method, which detects nucleic acid instead of protein, is a much more sensitive method compared to conventional PCR method, with shorter analytical time and lower detection limit. Its high specificity and sensitivity, together with its semiquantitative ability, give it a huge potential to serve as a powerful detection tool in various industries such as medical, veterinary, and agricultural industries. With the recent advances in PCR-ELISA, it is envisaged that the assay is more widely recognized for its fast and sensitive detection limit which could improve overall diagnostic time and quality.

scholarly journals Immunostaining of DNA in electron microscopy: an amplification and staining procedure for thin sections as alternative to gold labeling.

1994 ◽  
Vol 42 (5) ◽  
pp. 635-643 ◽  
Author(s):  
B Bohrmann ◽  
E Kellenberger
Keyword(s):  
Freeze Substitution ◽  
High Specificity ◽  
Staining Technique ◽  
Uranyl Acetate ◽  
Staining Procedure ◽  
Igg Antibody ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Specificity And Sensitivity ◽  
Eukaryotic Chromatin

We describe a new electron microscopic on-section staining technique with high specificity and sensitivity for DNA-containing structures. Lowicryl HM20 sections of specimens obtained by cryofixation and freeze-substitution are incubated in a first step with a primary IgM antibody specific for double-stranded DNA. The layer of bound antibodies at the section surface is amplified in a successive step by a secondary IgG antibody. Finally, electron scattering of the antibody layer produced is enhanced by staining with a mixture of uranyl acetate and potassium permanganate. The applicability of the method is exemplified by the detection of shape and distribution of various types of bacterial and eukaryotic chromatin.

Polymerase chain reaction amplification and typing of rotavirus in environmental samples

1995 ◽  
Vol 31 (5-6) ◽  
pp. 371-374 ◽  
Author(s):  
R. Gajardo ◽  
R. M. Pintó ◽  
A. Bosch
Keyword(s):  
Polymerase Chain Reaction ◽  
Environmental Samples ◽  
Polymerase Chain Reaction Amplification ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Group A ◽  
Reaction Amplification ◽  
Stool Specimens ◽  
Polymerase Chain

A reverse transcription polymerase chain reaction (RT-PCR) assay is described that has been developed for the detection and serotyping of group A rotavirus in stool specimens and concentrated and non-concentrated sewage specimens.

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