Conventional methods for H5 virus detection are time-consuming BAY 11-7082 clinical trial and technically demanding, and most importantly, these methods are not practical for field investigation [17]. Several rapid diagnostic kits for the detection of H5 subtype viruses have been reported. But more than a couple of monoclonal antibodies or polyclonal antibodies are required to reach appropriate specificity and sensitivity of detection, which increases production

cost [14]. The application of the complementary Mab pair reported in this study provides a solution to this and makes it possible for the cost effective production of rapid H5 tests for field usage. One of the H5 strains from chicken, which can not be detected by the dot ELISA, was subjected to HA sequencing.

The sequence result indicated that multiple deletions occur in its H5 sequence, such as the 353rd and the 387th nucleotide. These mutations may cause changes in HA protein structure and abolish the interaction to specific Mabs. These nature virus mutants may not replicate properly GW3965 ic50 and spread efficiently due to their genetic instability. Therefore, it is concluded that the dot ELISA performed here is able to detect those circulating H5N1 viruses that did not change genetically in their HA genes. Unlike chicken, duck and other water fowls do not show any symptom even if they are infected with high concentration of H5 virus [18]. These virus carriers can cause virus shedding and spreading. Virus titration N-acetylglucosamine-1-phosphate transferase studies indicate that these non-symptomatic birds can shed more than 108 EID50/ml of the virus to the environment. The dot test developed here is sensitive

enough to achieve specific early detection in poultry species. Therefore, the use of the H5 dot ELISA rapid test on site will reduce the risk of the false negative results via symptom observation only. Though, as a common challenge for all the rapid field tests, there is the possibility of false negative results due to the limitation of test sensitivity, this H5 dot ELISA serves as an effective tool for H5 screening at the very early stage. For those possible infected populations, it is still necessary to confirm with RT-PCR after the primary H5 infection screening with this rapid test first, if the clinical condition allows. Selection of the H5 HA specific MAbs for the development of the H5 dot ELISA was based on detailed analyses of their PF-3084014 binding properties.