scholarly journals Use of Peptide-Based Enzyme-Linked Immunosorbent Assay followed by Immunofluorescence Assay To Document Ehrlichia chaffeensis as a Cause of Febrile Illness in Nicaragua

2016 ◽  
Vol 54 (6) ◽  
pp. 1581-1585 ◽  
Author(s):  
Ijeuru Chikeka ◽  
Armando J. Matute ◽  
J. Stephen Dumler ◽  
Christopher W. Woods ◽  
Orlando Mayorga ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Large Scale ◽  
Febrile Illness ◽  
The United States ◽  
Immunofluorescence Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Ehrlichia Chaffeensis ◽  
Acute Febrile Illness ◽  
Serum Samples ◽  
Content Type

Ehrlichia chaffeensis, the etiologic agent of human monocytic ehrlichiosis (HME), has been extensively studied as a cause of acute febrile illness and an emerging tick-borne zoonosis in the United States. Limited data suggest its presence in other regions, including Central and South America but not Nicaragua to date. Diagnosis ofE. chaffeensisinfection by indirect immunofluorescence assay (IFA) is the reference standard due to its presumed high sensitivity and specificity, but IFA is impractical, variably reproducible, and cumbersome for large epidemiologic studies and for clinical diagnosis in resource-poor regions. We evaluated a high-throughput, objective peptide-based enzyme-linked immunosorbent assay (ELISA) for use alone or in combination with IFA. We found that it performed best as a screening test (sensitivity, 100%; specificity, 84%) to reduce the proportion of serum samples that were required by the more cumbersome and subjective IFA testing to <20%. Using a two-step diagnostic approach (IFA is performed if the ELISA is positive), we identifiedE. chaffeensisor a serologically and antigenically similar organism as a heretofore unrecognized cause of acute febrile illness in humans in Nicaragua and demonstrated the utility of the peptide ELISA as a screening tool for large-scale clinical studies.

scholarly journals Development of a Novel Cocktail Enzyme-Linked Immunosorbent Assay and a Field-Applicable Lateral-Flow Rapid Test for Diagnosis of Contagious Bovine Pleuropneumonia

2016 ◽  
Vol 54 (6) ◽  
pp. 1557-1565 ◽  
Author(s):  
Martin Heller ◽  
Nimmo Gicheru ◽  
Georgina Tjipura-Zaire ◽  
Cecilia Muriuki ◽  
Mingyan Yu ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Rapid Test ◽  
Lateral Flow ◽  
Sub Saharan Africa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Contagious Bovine Pleuropneumonia ◽  
Serum Samples ◽  
Content Type ◽  
Mycoplasma Mycoides ◽  
Sub Saharan

Contagious bovine pleuropneumonia (CBPP) is a severe respiratory disease that is widespread in sub-Saharan Africa. It is caused byMycoplasma mycoidessubsp.mycoides, a bacterium belonging to theMycoplasma mycoidescluster. In the absence of an efficient CBPP vaccine, improved and easy-to-use diagnostic assays for recurrent testing combined with isolation and treatment of positive animals represent an option for CBPP control in Africa. Here we describe the comprehensive screening of 17 immunogenicMycoplasma mycoidessubsp.mycoidesproteins using well-characterized bovine sera for the development of a novel cocktail enzyme-linked immunosorbent assay (ELISA) for laboratory use. Two recombinantMycoplasmaimmunogens, MSC_0136 and MSC_0636, were used to set up a standardized cocktail ELISA protocol. According to the results from more than 100 serum samples tested, the sensitivity and specificity of the novel cocktail ELISA were 85.6% and 96.4%, respectively, with an overall diagnostic accuracy comparable to that of the Office International des Epizooties (OIE)-prescribed serological assays. In addition, we provide a proof of principle for a field-applicable, easy-to-use commercially produced prototype lateral-flow test for rapid (<30-min) diagnosis of CBPP.

scholarly journals Detection of Francisella tularensis-Specific Antibodies in Patients with Tularemia by a Novel Competitive Enzyme-Linked Immunosorbent Assay

2012 ◽  
Vol 20 (1) ◽  
pp. 9-16 ◽  
Author(s):  
Neekun Sharma ◽  
Akitoyo Hotta ◽  
Yoshie Yamamoto ◽  
Osamu Fujita ◽  
Akihiko Uda ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Francisella Tularensis ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
The Novel ◽  
Serum Samples ◽  
Content Type ◽  
Microagglutination Test ◽  
Highly Sensitive ◽  
High Degree

ABSTRACTA novel competitive enzyme-linked immunosorbent assay (cELISA) was developed and evaluated for detection of antibodies againstFrancisella tularensisin humans. The assay is based on the ability of serum antibodies to inhibit the binding of monoclonal antibodies (MAbs) directed againstF. tularensislipopolysaccharide antigens. The assay was evaluated using serum samples of tularemia patients, inactivatedF. tularensis-immunized rabbits, andF. tularensis-infected mice. Antibodies againstF. tularensiswere successfully detected in serum samples of tularemia patients as well as the immunized and infected animals. The cELISA method was compared to indirect ELISA (iELISA) and the commonly used microagglutination test (MA) using serum samples of 19 tularemia patients and 50 healthy individuals. The sensitivity and specificity of cELISA were 93.9 and 96.1%, respectively, in comparison to the iELISA. MA was less sensitive than cELISA with a sensitivity and specificity of only 81.8 and 98.0%, respectively. A high degree of correlation (R2= 0.8226) was observed between cELISA and iELISA results. The novel cELISA developed in this study appears to be highly sensitive and specific for serodiagnosis of human tularemia. The potential of the MAb-based cELISA to be used in both human and animal samples emphasizes its usefulness for serological survey of tularemia among multiple animal species.

scholarly journals Development of an Enzyme-Linked Immunosorbent Assay for Serodiagnosis of Ringworm Infection in Cattle

2013 ◽  
Vol 20 (8) ◽  
pp. 1150-1154 ◽  
Author(s):  
Elena Tatiana Băguţ ◽  
Ludivine Cambier ◽  
Marie-Pierre Heinen ◽  
Vasile Cozma ◽  
Michel Monod ◽  
...  
Keyword(s):  
Positive Predictive Value ◽  
Negative Predictive Value ◽  
Immunosorbent Assay ◽  
Predictive Value ◽  
Skin Lesions ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Predictive Values ◽  
Content Type ◽  
Ringworm Infection

ABSTRACTThe aim of this study was to develop an in-house enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of ringworm infection in cattle. We used available recombinant forms ofTrichophyton rubrumdipeptidyl peptidase V (TruDppV) andT. rubrumleucin aminopeptidase 2 (TruLap2), which are 98% identical toTrichophyton verrucosumorthologues. Field serum samples from 135 cattle with ringworm infection, as confirmed by direct microscopy, fluorescence microscopy, and PCR, and from 55 cattle without any apparent skin lesions or history of ringworm infection that served as negative controls were used. Sensitivities, specificities, and positive and negative predictive values were determined to evaluate the diagnostic value of our ELISA. Overall, the ELISAs based on recombinant TruDppV and TruLap2 discriminated well between infected animals and healthy controls. Highly significant differences (P< 0.0001, Mann-Whitney U test) were noted between optical density values obtained when sera from infected versus control cattle were tested. The ELISA developed for the detection of specific antibodies against DppV gave 89.6% sensitivity, 92.7% specificity, a 96.8% positive predictive value, and a 78.4% negative predictive value. The recombinant TruLap2-based ELISA displayed 88.1% sensitivity, 90.9% specificity, a 95.9% positive predictive value, and a 75.7% negative predictive value. To the best of our knowledge, this is the first ELISA based on recombinant antigens for assessing immune responses to ringworm infection in cattle; it is particularly suitable for epidemiological studies and also for the evaluation of vaccines and/or vaccination procedures.

scholarly journals Sensitive and Specific Enzyme-Linked Immunosorbent Assay for Detecting Serum Antibodies against Mycobacterium avium subsp. paratuberculosis in Fallow Deer

2014 ◽  
Vol 21 (8) ◽  
pp. 1077-1085 ◽  
Author(s):  
José M. Prieto ◽  
Ana Balseiro ◽  
Rosa Casais ◽  
Naiara Abendaño ◽  
Liam E. Fitzgerald ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Mycobacterium Avium ◽  
Cost Effective ◽  
Fallow Deer ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Igg Antibody ◽  
Tissue Samples ◽  
Content Type ◽  
Mycobacterium Avium Subsp Paratuberculosis

ABSTRACTThe enzyme-linked immunosorbent assay (ELISA) is the diagnostic test most commonly used in efforts to control paratuberculosis in domestic ruminants. However, commercial ELISAs have not been validated for detecting antibodies againstMycobacterium aviumsubsp.paratuberculosisin wild animals. In this study, we compared the sensitivities and specificities of five ELISAs using individual serum samples collected from 41 fallow deer with or without histopathological lesions consistent with paratuberculosis. Two target antigenic preparations were selected, an ethanol-treated protoplasmic preparation obtained from a fallow deerM. aviumsubsp.paratuberculosisisolate (ELISAs A and B) and a paratuberculosis protoplasmic antigen (PPA3) (ELISAs C and D). Fallow deer antibodies bound to the immobilized antigens were detected by using a horseradish peroxidase (HRP)-conjugated anti-fallow deer IgG antibody (ELISAs A and C) or HRP-conjugated protein G (ELISAs B and D). A commercially available assay, ELISA-E, which was designed to detectM. aviumsubsp.paratuberculosisantibodies in cattle, sheep, and goats, was also tested. Although ELISAs A, C, and E had the same sensitivity (72%), ELISAs A and C were more specific (100%) for detecting fallow deer with lesions consistent with paratuberculosis at necropsy than was the ELISA-E (87.5%). In addition, the ELISA-A was particularly sensitive for detecting fallow deer in the latent stages of infection (62.5%). The antibody responses detected with the ELISA-A correlated with both the severity of enteric lesions and the presence of acid-fast bacteria in gut tissue samples. In summary, our study shows that the ELISA-A can be a cost-effective diagnostic tool for preventing the spread of paratuberculosis among fallow deer populations.

scholarly journals Scrub typhus in patients with acute febrile illness: a 5-year study from India

QJM ◽  
2019 ◽  
Vol 113 (6) ◽  
pp. 404-410
Author(s):  
C K Thakur ◽  
R Chaudhry ◽  
N Gupta ◽  
E V Vinayaraj ◽  
V Singh ◽  
...  
Keyword(s):  
Scrub Typhus ◽  
Monsoon Season ◽  
Geographical Location ◽  
Febrile Illness ◽  
Immunofluorescence Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Acute Febrile Illness ◽  
Nested Polymerase Chain Reaction ◽  
Pulmonary Manifestations ◽  
Post Monsoon Season

Abstract Background Scrub typhus was once thought to be a disease of rural origin and was confined to specific pockets in South Asia. Early diagnosis and treatment is extremely important as it is associated with high mortality if left untreated. Aim To delineate the clinical and molecular epidemiology of scrub typhus in patients presenting with acute febrile illness from various parts of India. Methods During the study period of 5 years (October 2013 to October 2018), a total of 1742 patients with acute febrile illness &lt;15 days were enrolled after taking informed consent. Patients were diagnosed using IgM Enzyme-linked immunosorbent assay (ELISA) based on the pre-determined region specific cut offs. Patients with positive IgM ELISA were also subjected to IgM Immunofluorescence assay and nested polymerase chain reaction (PCR) assay. The demographic and relevant clinical details of the patients were documented and analyzed. Results A total of 210 (12.1%) patients were diagnosed with scrub typhus. Of these, nested PCR was positive in only 85 patients. Sequencing and phylogenetic analysis showed that the predominant circulating genotypes were Gilliam and Karp. On multivariate analysis, acute respiratory distress syndrome, myocarditis, encephalitis/encephalopathy, jaundice and splenomegaly were significantly more common in those patients who were diagnosed with scrub typhus. A total of 14 patients diagnosed with scrub typhus succumbed to the illness. Conclusion Patients with fever, headache, pulmonary manifestations, CNS manifestations, myocarditis, transaminitis or thrombocytopenia presenting in the monsoon and post-monsoon season should be evaluated for scrub typhus irrespective of the geographical location in India.

scholarly journals Sunbathing, a possible risk factor of murine typhus infection in Greece

2021 ◽  
Vol 15 (3) ◽  
pp. e0009186
Author(s):  
Stavroula Labropoulou ◽  
Ekatherina Charvalos ◽  
Stylianos Chatzipanagiotou ◽  
Anastasios Ioannidis ◽  
Panagiotis Sylignakis ◽  
...  
Keyword(s):  
Large Scale ◽  
Febrile Illness ◽  
Epidemiological Data ◽  
Immunofluorescence Assay ◽  
Murine Typhus ◽  
Serum Samples ◽  
Epidemiological Features ◽  
Severe Infections ◽  
Classical Triad ◽  
Of Greece

Background There are few studies about the presence of murine typhus in Greece. Our objective was to conduct a large scale retrospective investigation to determine the clinical and epidemiological features of patients diagnosed with murine typhus in Greece. Methodology/Principal findings From 2012 to 2019 serum samples from hospitalized patients and outpatients throughout Greece suspected for murine typhus infection were tested by immunofluorescence assay for Rickettsia typhi. Immunofluorescence positive samples obtained since 2016 were also tested by qPCR targeting R. typhi. Clinical and epidemiological data were retrospectively collected for the patients with confirmed murine typhus. Overall, we tested 5,365 different patients and, in total, 174 patients from all geographic regions of Greece were diagnosed with murine typhus. The most frequently reported sign or symptom was fever (89%), followed by headache (84%) and rash (81%). The classical triad of fever, headache, and rash was present in 72% of patients during their illness. Severe infections with complications including acute renal failure or septic shock were not recorded. The majority of cases (81%) occurred during May–October and peaked in June and September. Most of patients (81%) infected in Athens, recalled that their only activity the last weeks before symptoms onset was swimming on the beach and 59% of them also reported an insect bite while sunbathing. Conclusions/Significance Our results may reflect the reemergence of murine typhus in Greece and we highlight the importance of awareness of this difficult-to-recognize undifferentiated febrile illness.

scholarly journals An enzyme-linked immunosorbent assay for the detection of mouse polyomavirus-specific antibodies in laboratory mice

1994 ◽  
Vol 28 (3) ◽  
pp. 257-261 ◽  
Author(s):  
H. W. J. Broeders ◽  
J. Groen ◽  
G. van Steenis ◽  
A. D. M. E. Osterhaus
Keyword(s):  
Immunosorbent Assay ◽  
Haemagglutination Inhibition ◽  
Immunofluorescence Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Pathogen Free ◽  
Laboratory Mice ◽  
Serum Samples ◽  
Laboratory Rodents ◽  
Antibody Titres ◽  
Specific Pathogen

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection and quantification of IgM and IgG serum antibodies to mouse polyomavirus (MPV). To evaluate the potential of this ELISA for the screening of laboratory rodents, serum samples from specific pathogen free (SPF) BALB/c RIVM mice, collected after experimental intraperitoneal infection with MPV, were tested by this assay. The results were compared with those obtained from the same sera in an immunofluorescence assay (IFA) and a haemagglutination inhibition assay (HIA). The ELISA proved to be the most sensitive of the 3 assays, allowing the detection of seropositive animals within 7 days post-infection and giving antibody titres that were about 4 to 8 times higher than those found in the IFA and HIA respectively. More than 5000 serum samples from non-infected specific pathogen free laboratory mice and 90 from 10 SPF N:NIH/RIVM mice experimentally infected with K-papovavirus, were negative in this assay, thus confirming the specificity of the ELISA

scholarly journals Recombinant Antigen-Based Enzyme-Linked Immunosorbent Assay for Diagnosis of Baylisascaris procyonis Larva Migrans

2011 ◽  
Vol 18 (10) ◽  
pp. 1650-1655 ◽  
Author(s):  
Sriveny Dangoudoubiyam ◽  
Ramesh Vemulapalli ◽  
Momar Ndao ◽  
Kevin R. Kazacos
Keyword(s):  
Immunosorbent Assay ◽  
Cross Reactivity ◽  
Larva Migrans ◽  
Enzyme Linked Immunosorbent Assay ◽  
Parasitic Infections ◽  
Serum Samples ◽  
Western Blot Assay ◽  
Human Patient ◽  
Content Type ◽  
Baylisascaris Procyonis

ABSTRACTBaylisascarislarva migrans is an important zoonotic disease caused byBaylisascaris procyonis, the raccoon roundworm, and is being increasingly considered in the differential diagnosis of eosinophilic meningoencephalitis in children and young adults. Although aB. procyonisexcretory-secretory (BPES) antigen-based enzyme-linked immunosorbent assay (ELISA) and a Western blot assay are useful in the immunodiagnosis of this infection, cross-reactivity remains a major problem. Recently, a recombinantB. procyonisantigen, BpRAG1, was reported for use in the development of improved serological assays for the diagnosis ofBaylisascarislarva migrans. In this study, we tested a total of 384 human patient serum samples in a BpRAG1 ELISA, including samples from 20 patients with clinicalBaylisascarislarva migrans, 137 patients with other parasitic infections (8 helminth and 4 protozoan), and 227 individuals with unknown/suspected parasitic infections. A sensitivity of 85% and a specificity of 86.9% were observed with the BpRAG1 ELISA, compared to only 39.4% specificity with the BPES ELISA. In addition, the BpRAG1 ELISA had a low degree of cross-reactivity with antibodies toToxocarainfection (25%), while the BPES antigen showed 90.6% cross-reactivity. Based on these results, the BpRAG1 antigen has a high degree of sensitivity and specificity and should be very useful and reliable in the diagnosis and seroepidemiology ofBaylisascarislarva migrans by ELISA.

scholarly journals Comparison of Enzyme-Linked Immunosorbent Assay, Western Blotting, Microagglutination, Indirect Immunofluorescence Assay, and Flow Cytometry for Serological Diagnosis of Tularemia

2004 ◽  
Vol 11 (6) ◽  
pp. 1008-1015 ◽  
Author(s):  
Mustafa Porsch-Özcürümez ◽  
Nele Kischel ◽  
Heidi Priebe ◽  
Wolf Splettstösser ◽  
Ernst-Jürgen Finke ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Western Blotting ◽  
Francisella Tularensis ◽  
Immunosorbent Assay ◽  
Indirect Immunofluorescence ◽  
Immunofluorescence Assay ◽  
Indirect Immunofluorescence Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Diagnostic Sensitivity And Specificity

ABSTRACT The serodiagnostic efficiencies of five different approaches to detecting antibodies (immunoglobulins G, A, and M) developed in clinically proven infections with Francisella tularensis have been assessed. Fifty serum samples from patients suffering from tularemia during an outbreak in Sweden were compared with samples from 50 healthy blood donors (controls) by using an enzyme-linked immunosorbent assay (ELISA), microagglutination (MA), Western blotting (WB), an indirect immunofluorescence assay (IIFA), and flow cytometry (FC). ELISA, WB, and FC were based on the use of preparations of lipopolysaccharides (LPS) of the live vaccine strain of Francisella tularensis subsp. holarctica (ATCC 29684) as a capture antigen. Whole methanol-fixed bacteria were used for IIFA and MA. Optimized protocols yielded a diagnostic sensitivity and specificity of 100% for WB, MA, and FC, 98% for ELISA, and 93% for IIFA. A total of 6,632 serum samples from individuals between the ages of 18 and 79 years, representatively recruited from all regions of Germany, were screened to estimate and confirm the positive predictive value (PVpos) of the ELISA. Serum samples from 15 (0.226%) individuals tested positive for F. tularensis-specific antibodies by ELISA and confirmatory WB. The resulting prevalence-dependent PVpos of 10.2% and specificity of 98.1% were consistent with our findings for tularemia patients and controls. We conclude that the combined usage of a screening ELISA and a confirmatory WB based on LPS as a common antigen, as well as the MA, is a suitable serodiagnostic tool, while the quality of the IIFA is hampered by subjective variations of the results. FC is a promising new approach that might be improved further in terms of multiplex analyses or high-throughput applications.

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