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Journal of Infection and Molecular Biology

Research Article
J. Inf. Mol. Biol. 2 (1): 1 - 5
http://dx.doi.org/10.14737/jimb.2307-5465/2.1.1.5
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Shahid Khan¹*, Shamsher Ali Khan¹, Sayyed Rahimullah Shah¹, Faizul Hassan¹, Kaleem Allah¹, Khasro Kaleem²
¹Microbiology Section, Veterinary Research and Disease Investigation Centre, Swat, KPK province, Pakistan.
²Livestock Research and Development Station Dir (Lower), KPK province, Pakistan
*Corresponding author: wazirkhattak@yahoo.com

ABSTRACT
Preparation of autogenous vaccine against Hydropericardium syndrome (HPS) and subsequent administration to chicks is a common practice in Pakistan. However, due to lack of quality control testing and evaluation procedures, the accurate quantity of virus from a given clinical sample of liver always remains questionable. Therefore, the present study has been conducted to differentiate the protein profile of normal and HPS infected liver lysate with respect to the availability of immunogenic proteins using sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and western blotting techniques. Using commercially available vaccine against HPS (Ottomans Parma), hyperimmune serum was raised in immunologically naïve broiler birds (3–weeks old, n = 60) and subjected to agar gel precipitation test (AGPT) for screening HPS positive serum samples. Of the total examined birds, only five birds were found positive. The commercially available HPS autogenous vaccine, liver from HPS positive bird and normal liver samples were subjected to protein extraction and measurement. SDS–PAGE analysis revealed 15–structural polypeptide bands in both HPS infected poultry liver and autogenous vaccine. Contrary to this, 10–structural polypeptide bands were observed in normal liver. The polypeptides of HPS autogenous vaccine and both normal and HPS infected livers, separated on 9% gel were transferred onto nitrocellulose membrane. The western blot analysis of proteins from Hydropericardium virus infected liver and HPS autogenous vaccine showed one immunogenic protein of molecular weight 15-20 kDa These differences could be used for identification and differentiation between clinically diseased and healthy birds as well as vaccinated by autogenous vaccine.

Key Words: Hydropericardium syndrome (HPS), AGPT, Sodium dodecyle sulphate polyacrylamide gel electrophoresis (SDS–PAGE), Western blotting technique