Abstract
<p class="abstrak2">Rapid diagnostic tools or point-of-care (POC) test is needed in the effort to control and eradicate the high pathogenic avian influenza (HPAI) H5N1 in Indonesia. Accuracy of a POC test is determined by the specificity of antibodies, which is the main component of a POC test. Recently a linear epitope, CNTCKQTP epitope, located at 274-281 amino acid residue of H5 hemagglutinin has been confirmed to be present all clade of H5N1 viruses. This study aimed at producing and evaluating the reactivity of a monospecific, polyclonal antibody against the epitope. The Antibody was produced by immunising a goat with the peptide in the form of multiple antigen peptide (MAP). The specificity of the antibody was estimated by assaying its reactivity against influenza virus subtypes H3N3, H4N4, H5N1, H6N5, H7N7, H9N2, H10N7 and H11N9; and recombinant hemagglutinins H1-H12, H14 and H15 with ELISA and immunoblot. The results of the assay showed that CNTKCQTP antibody was not specific for H5 haemagglutinin because it cross-reacted with other haemagglutinins especially H7, H8 and H9. The potential of the peptide containing the epitope, GNCNTKCQTPMGAINSS. as an ELISA reagent for assaying H5 antibodies in chickens previously vaccinated and challenged with the H5N1 virus was also evaluated in this study. In contrast, the results of previous studies, the ELISA using GNCNTKCQTPMGAINSS as coating antigen was not sensitive in detecting antibody to haemagglutinin H5 in chickens.</p><strong>Key Words: </strong>AI Virus, Hemagglutinin H5, CNTKCQTP Epitope, MAP, Immunoassa<em>y</em>
Highlights
In the early 2000s, highly pathogenic avian influenza (HPAI) H5N1 influenza was detected in many parts of the world causing concerns after eighteen people were hospitalized, six of whom died, after contracting the H5N1 virus from infected chickens (Claas et al 1998)
After four vaccinations the serum had a high titre of antibodies as it recognised the CNTKCQTP multiple antigen peptide (MAP) at high 1 : 125,600 dilution
The titre with GNCNTKCQTPMGAINSS single peptide was much lower than the CNTKCQTP MAP
Summary
In the early 2000s, highly pathogenic avian influenza (HPAI) H5N1 influenza was detected in many parts of the world causing concerns after eighteen people were hospitalized, six of whom died, after contracting the H5N1 virus from infected chickens (Claas et al 1998). Most of the 63 affected countries were able to eliminate H5N1 virus. One of the most important factors for those countries that successfully eradicated H5N1 virus was their ability to quickly recognise the disease enabling rapid elimination of all infected birds. To be able to recognize a disease quickly, the availability of appropriate diagnostic techniques is of paramount importance. For countries where the H5N1 has became endemic, two types of diagnostic tests are necessary; the first type that includes virus isolation and real time PCR, is intended for confirmatory, unambiguous diagnosis, whereas the second type is the point-of-care, or pen-site test intended for rapid diagnosis and to guide in determining actions needed to be taken for control or eradication of disease (Tarigan 2015). Most tests of the second type are immunological test based on the use of an antibody to detect the presence of H5N1 antigen
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