Identification and Titration of Hydro Pericardium Syndrome Virus (HPSV) by using Modified Counter-Current Immuno-Electrophoresis (MCCIE)

Abstract

A rapid, simple, and accurate Modified Counter Current Immuno-electrophoresis (MCCIE) technique was developed and compared with the Reverse Passive Heamagglutination Assay (RPHA) for the titration of HPS virus. The MCCIE test had 100% correlation with the conventional RPHA and correctly titrated 116 samples which were already titrated with RPHA. Instead of horizontal electrophoresis chamber used in counter current Immuno-electrophoresis test, a modification in Counter Current Immuno-Electrophoresis (CCIE) test was made by using 1% melted agarose gel filled in 8 cm long and 3 mm narrow U-shaped glass tubing for the detection as well as quantification & titration of HPS virus. Results of Liver samples of 116 poultry birds including 16 broilers (6 livers were from clinically positive), 50 Desi and 50 commercial layers (adult) obtained through modified counter current Immuno-electrophoresis (MCCIE) were found similar to those found though CCIE and titres obtained through MCCIE were found similar to RPHA tests. The titres in MCCIE test were expressed as appearance of hazy colored precipitation band in the twelve U-shaped tubes. All of the six clinically positive liver samples gave positive results with titres ranging from 1:32 to 1:128. Of these 6 positive, 3 samples had 1:32, 2 samples had 1:64 and 1 sample had 1:128 titres. While none of desi and commercial layer was found to be positive. The results indicated that MCCIE is simple and inexpensive test as compared to CCIE & RPHA to detect and titrate HPS virus in the infected poultry liver samples. All the HPS virus isolates from HPS infected broiler livers and adenovirus group-I obtained from NARC, Islamabad and VRI, Lahore showed MCCIE & RPHA titre (1:64) with specific HPS antibodies.