Rapid diagnosis of influenza infection of NP antigen using an immunocapture ELISA test

Chomel, Jean-Jacques; Thouvenot, D.; Onno, M.; Kaiser, C.; Gourreau, Jean-Marie; Aymard, M.

doi:10.1016/0166-0934(89)90102-x

Cited by 50 publications

(28 citation statements)

References 12 publications

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“…In another aliquot of the nasal swab, we searched for antigens with an Elisa immunocapture test [10,11]. For influenza A, we used the monoclonal antibody NPA 153 kindly supplied by A. Douglas (W.I.C., London, UK) for influenza B, a monoclonal antibody (8F1OE10) produced in house, lor the parainfluenza viruses 1 + 2 + 3. a supernatant of hybridoma cell culture (GF8D) produced in house, and the respective polyclonal antisera prepared in rabbits immunized with purified concentrated viruses mixed with complete Freund's adjuvant.…”

Section: Methodsmentioning

confidence: 99%

Epidemiology of Viral Infections and Evaluation of the Potential Benefit of OM-85 BV on the Virologie Status of Children Attending Day-Care Centers

Aymard¹,

Chomel²,

Allard³

et al. 1994

Respiration

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Viral investigations were performed during 4 winter seasons (88/89, 89/90, 92/93, 93/94) in children attending day-care centers (DCCs) in the Rhone Département in eastern France. Over the total observation period of 4 winter seasons, 780 children were screened with a nasal swab for the presence of viruses. Of those, 230 (29.5%) had a positive viral culture. The viruses identified were respiratory syncytial virus (RSV), influenza A and B virus, parain-fluenza virus, coronavirus, rhinovirus, adenovirus and enterovirus. During that time, 83 epidemic events in 47 DCC were recorded. A particular virus was judged to be causally related to an epidemic if the identical virus was isolated in ≥ 3 children during the same outbreak of respiratory diseases. Thus, in 51 cases (61.4%) of all epidemics, the following viruses were responsible for an epidemic: RSV (n = 23), coronavirus (n = 10) (only during the season of 1993-1994), influenza A virus (n = 6), rhinovirus (n = 4), enterovirus (n = 4), adenovirus (n = 3) and parainfluenza virus (n = 1). Except for the somewhat surprising accumulation of coronavirus epidemics during the winter of 1993-1994, there were only minor seasonal variations from one year to another. As expected, RSV accounted for about one third of all respiratory tract infections in children attending DCCs and was therefore the most important single causative agent. These results are compared with data from children who did not attend a DCC and were cared for in a private practice. During the winter of 1989-1990, the viral epidemiological survey was performed at the same time and in parallel to a double-blind, placebo-controlled clinical study investigating the efficacy of OM-85 BV, an immunoactive bacterial extract. This study, enrolling 423 children attending DCCs demonstrated a protective effect of OM-85 BV in significantly reducing the risk of recurrent infections of the upper respiratory tract during the treatment period with the compound. 34% of all participating children (75 in the verum group, 70 in the placebo group) were enrolled in an additional virological study. In these patients, RSV was isolated 10 times in the placebo group, but only 5 times in the treated group (p < 0.05) and influenza A virus was present in 4 children in the placebo group, but only in 1 infant in the verum group giving a total of 14 positive virologie results in the placebo group versus 6 in the verum group (p < 0.05). Despite the small numbers of children investigated for their virologie status during respiratory infectious outbreaks, there was a statistically significant difference in the prevalence of virus carriers in favor of the children treated with OM-85 BV. These results corroborate the clinical findings.

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Section: Methodsmentioning

confidence: 99%

Epidemiology of Viral Infections and Evaluation of the Potential Benefit of OM-85 BV on the Virologie Status of Children Attending Day-Care Centers

Aymard¹,

Chomel²,

Allard³

et al. 1994

Respiration

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“…The same was true with field sera, in which HI detected antibody to a single subtype and IPMA detected antibodies to both subtypes (Table 2). This is not surprising because pigs develop antibody to both internal and external proteins of SIV after natural infection 3 and IPMA detects antibodies regardless of the hemagglutinin subtype used. The ability of the HI test to detect antibody against a particular strain of SIV, on the other hand, depends on the antigenic relationship between the virus producing the antibody and the virus isolate used in the test.…”

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confidence: 99%

An Immunoperoxidase Monolayer Assay for the Detection of Antibodies against Swine Influenza Virus

2002

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Abstract. An immunoperoxidase monolayer assay (IPMA) has been developed to detect antibodies against swine influenza A virus (SIV) in pig sera. The test was evaluated by using sequential sera from pigs experimentally infected with H1N1 subtype of SIV. Two hundred field serum samples that had been examined by the hemagglutination inhibition (HI) test were also tested. Antibodies specific to SIV were detected as early as 3 days postinoculation (dpi) in the IPMA test as compared with 7 dpi by the HI test. Unlike HI, no serum treatment was required in the IPMA test. Regardless of the virus used in the test, IPMA detected antibodies to both H1N1 and H3N2 subtypes of SIV whereas HI detects antibodies against either H1N1 or H3N2, depending upon the virus used in the test. Results of this study indicate that IPMA is a useful test for screening of pig sera for SIV antibodies.Swine influenza virus (SIV) belongs to the family Orthomyxoviridae. 4 There are three types of this virus based on the antigenic relatedness of nucleoprotein (NP) and matrix (M) proteins: A, B, and C. 12 Most of the swine influenza worldwide is caused by type A, 6 which is further divided into subtypes on the basis of its hemagglutinin (HA) and neuraminidase (N) surface proteins. The predominant subtypes reported to be associated with swine influenza throughout the world are H1N1, H1N2, and H3N2. 1,19 H1N1 has been the predominant subtype circulating in pigs in the United States. 2,5,14 Recently, however, H3N2 has been recovered from swine with acute clinical disease in the eastern and north central regions of the United States. 8,18,21,22 Subtype H1N2 has also been isolated recently from pigs in Indiana. 7 Serological diagnosis of SIV is currently based on hemagglutination inhibition (HI) tests. Since there is no crossreaction between hemagglutinins, 2 different HI tests are used to detect antibodies against H1N1 and H3N2 subtypes of SIV. A serological test that can detect anti-NP antibody would be useful in diagnosing infection with all type A influenza viruses because NP protein is relatively conserved among various SIV strains. 19 Sensitive enzyme-linked immunosorbent assays (ELISA) have also been developed for SIV. 11,16,20 However, substantial amounts of purified antigens 15 or recombinant protein are needed for coating ELISA plates. 16,20 The purpose of this study was to develop an immunoperoxidase monolayer assay (IPMA) for the detection of SIV type A antibodies and compare it to the HI tests.Two subtypes of SIV (H1N1 and H3N2) were used in both IPMA and HI tests. A/Sw/NJ/1979/H1N1 was obtained from the National Veterinary Services Laboratories, Ames, Iowa, and A/Sw/MN/1998/H3N2 was isolated from a field sample submitted to the Minnesota Veterinary Diagnostic Laboratory (MDVL). A field isolate, A/Sw/MN/1997/H1N1, recovered from a grower pig in Minnesota was used for ex- perimental infection of pigs. Each isolate was confirmed to be H1-or H3-type by previously described method. 9 Experimental sera were produced in 5-week-old pigs. Eleven pigs ...

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“…4,7,8,13,14,23 These assays were comparable in sensitivity to conventional egg inoculation procedures but required far less effort and processing time for completion. The virus-capture ELISA described in this study exhibited greater sensitivity (0.1 ng/assay well) than the previously described formats (0.8-2 ng/assay well), 13,14,23 possibly because of our use of a biotin-avidin signal amplification system.…”

Section: Discussionmentioning

confidence: 93%

“…28 Antigen-capture type ELISA formats have been described. 4,7,8,13,14,23 These assays were capable of detecting influenza virus at the 10 ng/ml concentration level. Of the techniques described here, these assays provide the most convenient mechanism for multiple sampling and an adequate level of sensitivity.…”

mentioning

confidence: 99%

ELISA Method for Detection of Influenza A Infection in Swine

et al. 1993

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Abstract. An antigen-capture enzyme-linked immunosorbent assay (ELISA) was developed to monitor virus shedding associated with experimental infection with a field strain of swine influenza in pigs. The assay consisted of a monoclonal anti-nucleoprotein capture antibody and a biotinylated rabbit anti-influenza A (H1N1) sandwich antibody. The antigen-capture system was capable of detecting as little as 1 ng/ml purified virus. The ELISA system surpassed egg cultivation procedures in the detection of low levels of shedding virus. Egg cultivation procedures indicated that most viral shedding had ceased by day 10 postinfection. In contrast, antigen-capture ELISA still showed an ongoing presence of viral antigen. A virus-capture ELISA, using this capture-sandwich antibody system, is equivalent in sensitivity to conventional egg inoculation procedures for the detection of the early phases of virus shedding. The automative potential of an ELISA-based system coupled with a substantially reduced assay time requirement give this virus-capture ELISA a distinct advantage over other cell culture or egg-based diagnostic techniques.

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Rapid diagnosis of influenza infection of NP antigen using an immunocapture ELISA test

Cited by 50 publications

References 12 publications

Epidemiology of Viral Infections and Evaluation of the Potential Benefit of OM-85 BV on the Virologie Status of Children Attending Day-Care Centers

Epidemiology of Viral Infections and Evaluation of the Potential Benefit of OM-85 BV on the Virologie Status of Children Attending Day-Care Centers

An Immunoperoxidase Monolayer Assay for the Detection of Antibodies against Swine Influenza Virus

ELISA Method for Detection of Influenza A Infection in Swine

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