Detection of immunoglobulin M (IgM), IgA, and IgG Norwalk virus-specific antibodies by indirect enzyme-linked immunosorbent assay with baculovirus-expressed Norwalk virus capsid antigen in adult volunteers challenged with Norwalk virus.

Canine distemper morbillivirus (CDV) infection causes a frequently fatal systemic disease in a broad range of carnivore species, including domestic dogs. In CDV infection, classical serology provides data of diagnostic and prognostic values (kinetics of seroconversion) and is also used to predict the optimal vaccination age of pups. Routine CDV serology is still based on time- and cost-intensive virus neutralization assays (V-NA). Here, we describe a new capture-sandwich enzyme-linked immunosorbent assay (ELISA) that uses recombinant baculovirus-expressed nucleocapsid (N) protein of a recent CDV wild-type isolate (2544/Han95) for the detection of CDV-specific antibodies in canine sera. Recombinant antigen was produced with high efficacy in Heliothis virescens larvae. The capture-sandwich ELISA enabled a clear-cut qualitative evaluation of the CDV-specific immunoglobulin G (IgG) and IgM serostatuses of 196 and 35 dog sera, respectively. Inter-rater agreement analysis (κ = 0.988) indicated that the ELISA can be used unrestrictedly as a substitute for the V-NA for the qualitative determination of CDV-specific IgG serostatus. In an attempt to semiquantify N-specific antibodies, a one-step-dilution (alpha method) IgG-specific ELISA was implemented. Alpha values of ≥50% showed very good inter-rater agreement (κ = 0.968) with V-NA titers of ≥1/100 50% neutralizing dose (ND50) as measured against the central European CDV wild-type isolate 2544/Han95 in canine sera originating from northern Germany. An ND50 titer of 1/100 is considered a threshold, and titers of ≥1/100 indicate a resilient, protective immunity. CDV N-specific antibodies of the IgM class were detected by the newly developed ELISA in 9 of 15 sera obtained from dogs with symptoms of acute distemper. In leucocytes of 5 of the 15 dogs (all of which were also IgM positive) CDV RNA was detected by reverse transcription (RT)-PCR. The recombinant capture-sandwich ELISA detecting N-specific antibodies of the IgG class provided superior sensitivity and specificity and thus represents a rapid and cost-effective alternative to classical CDV V-NA. By detection of specific IgM antibodies, the ELISA will be complementary to RT-PCR and V-NA in the diagnosis of acute distemper infections.

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Immunoglobulin M Antibody Test To Detect Genogroup II Norwalk-Like Virus Infection

Journal of Clinical Microbiology ◽

10.1128/jcm.37.9.2983-2986.1999 ◽

1999 ◽

Vol 37 (9) ◽

pp. 2983-2986 ◽

Cited By ~ 13

Author(s):

James P. Brinker ◽

Neil R. Blacklow ◽

Xi Jiang ◽

Mary K. Estes ◽

Christine L. Moe ◽

...

Keyword(s):

Monoclonal Antibody ◽

Virus Infection ◽

Antibody Test ◽

Virus Antigen ◽

Immunoglobulin M ◽

Enzyme Linked Immunosorbent Assay ◽

Norwalk Virus ◽

Viral Etiology ◽

Specific Immunoglobulin ◽

Adult Volunteers

Sera obtained from adult volunteers inoculated with genogroup II Norwalk-like viruses (NLVs), Hawaii virus, and Snow Mountain virus and from patients involved in outbreaks of gastroenteritis were tested for genogroup II NLV Mexico virus-specific immunoglobulin M (IgM) by use of a monoclonal antibody, recombinant Mexico virus antigen (rMXV)-based IgM capture enzyme-linked immunosorbent assay (ELISA). Sera from genogroup I Norwalk virus (NV)-inoculated volunteers and from patients involved in a genogroup I NLV outbreak were also tested. In sera from those infected with genogroup I NV or NLVs in volunteer and outbreak studies, only 3 of 25 were rMXV IgM positive; in contrast, 24 of 25 were IgM positive for recombinant NV (rNV). In sera from those infected with genogroup II NLVs in volunteer and outbreak studies, 28 of 47 were rMXV IgM positive and none were IgM positive for rNV, showing the specificity of each IgM test for its respective genogroup. In an outbreak of gastroenteritis not characterized as being of viral etiology but suspected to be due to NV, 7 of 13 persons had IgM responses to rMXV, whereas none had IgM responses to rNV, thus establishing the diagnosis as genogroup II NLV infection. The rMXV-based IgM capture ELISA developed is specific for the diagnosis of genogroup II NLV infections.

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Limited Diagnostic Capacities of Two Commercial Assays for the Detection of Leptospira Immunoglobulin M Antibodies in Laos

Clinical and Vaccine Immunology ◽

10.1128/cvi.00219-06 ◽

2006 ◽

Vol 13 (10) ◽

pp. 1166-1169 ◽

Cited By ~ 33

Author(s):

Stuart D. Blacksell ◽

Lee Smythe ◽

Rattanaphone Phetsouvanh ◽

Michael Dohnt ◽

Rudy Hartskeerl ◽

...

Keyword(s):

Diagnostic Accuracy ◽

Immunosorbent Assay ◽

Gold Standard ◽

Agglutination Test ◽

Immunoglobulin M ◽

Microscopic Agglutination Test ◽

Enzyme Linked Immunosorbent Assay ◽

Diagnostic Utility

ABSTRACT The diagnostic utility of immunochromatographic (Leptotek) and enzyme-linked immunosorbent assay (ELISA; Panbio) tests for the detection of Leptospira immunoglobulin M antibodies was assessed in febrile adults admitted in Vientiane, Laos. Both tests demonstrated poor diagnostic accuracy using admission serum (Leptotek sensitivity of 47.3% and specificity of 75.5%: ELISA sensitivity of 60.9% and specificity of 65.6%) compared to the Leptospira “gold standard” microscopic agglutination test.

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Development of an enzyme-linked immunosorbent assay for quantification of Salmonella enteritidis-specific antibodies in egg yolk

Brazilian Journal of Poultry Science ◽

10.1590/s1516-635x2006000400009 ◽

2006 ◽

Vol 8 (4) ◽

pp. 257-263

Author(s):

IT Tayeb ◽

P Nehme ◽

L Jaber ◽

EK Barbour

Keyword(s):

Immunosorbent Assay ◽

Salmonella Enteritidis ◽

Egg Yolk ◽

Enzyme Linked Immunosorbent Assay ◽

Specific Antibodies

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Analysis of Specific Immunoglobulin G Subclass Antibodies for Serological Diagnosis of Echinococcosis by a Standard Enzyme-Linked Immunosorbent Assay

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.5.5.613-616.1998 ◽

1998 ◽

Vol 5 (5) ◽

pp. 613-616 ◽

Cited By ~ 21

Author(s):

Felix Grimm ◽

Friedrich E. Maly ◽

Jian Lü ◽

Roberto Llano

Keyword(s):

Healthy Volunteers ◽

Immunosorbent Assay ◽

Immunoglobulin G ◽

Serological Diagnosis ◽

Enzyme Linked Immunosorbent Assay ◽

Parasitic Infections ◽

Igg Subclass ◽

Specific Antibodies ◽

Specific Igg4 ◽

Antibody Levels

ABSTRACT The potential roles of specific antibodies of the different immunoglobulin G (IgG) subclasses in the serological diagnosis of cystic echinococcosis (CE) and alveolar echinococcosis (AE) were investigated by an enzyme-linked immunosorbent assay based on hydatid fluid as antigen. Specific antibodies of subclass 1 were found to be of major importance. In sera collected at the time of diagnosis (i.e., before any therapeutic intervention was initiated) they could be demonstrated in 14 of 15 sera from patients with CE and in all 12 sera from patients with AE. The most discriminatory and the most specific antibodies found in this study belonged to IgG subclass 4. Only one false-positive reaction was observed with 253 sera from healthy volunteers, and no cross-reactions occurred in 80 sera from patients with different parasitic infections. Specific IgG4 antibodies could be demonstrated in 61.0 to 66.7% (CE) or 47.6 to 66.7% (AE) of the cases. Antibody levels of IgG subclass 2 were elevated only moderately, and subclass 3 antibodies were detected in a few cases only. In addition, nonspecific reactions in sera of healthy volunteers or patients with other parasitic infections could partially be attributed to antibodies of subclasses 2 and 3.

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