Detection of Duck Plague Virus by Reverse Passive Hemagglutination Test

Abstract

A reverse passive hemagglutination (RPHA) test was developed to detect duck plague virus (DPV). The technique used sheep erythrocytes stabilized with formaldehyde and pyruvaldehyde and coated with immunoglobulin G (IgG) containing anti-DPV antibody prepared from antiserum produced in sheep. Optimum coating of stabilized erythrocytes occurred at 25 C and pH 4.0 with a concentration of IgG of 20-40 micrograms/ml and a 90-min incubation period. The coated cells were stable for 40 days when stored at 4 C or for at least 4 months (the longest period tested) when frozen at -70 C or -196 C. The RPHA test was conducted at 25 C and read after 3 hours. The high specificity of the test is indicated by the absence of cross-reactions with heterologous virus strains, with specimens prepared from normal duck livers, and with normal chicken embryo chorioallantoic fluid, as well as by the inhibition of hemagglutination only with DPV antiserum. The RPHA test detected six strains of DPV in all virus-containing specimens as well as the immunofluorescence (IF) test did; however, conventional plaque assays (PA) failed to detect virus in five specimens that contained three non-plaque-forming strains of DPV. The mean quantity of DPV that could be detected in the RPHA test was 25 plaque-forming units or 65 fluorescent units per ml. Although the RPHA test was less sensitive than either the PA or the IF test, there was a positive correlation in the titers of DPV antigens between all three tests. The RPHA test is a rapid, simple procedure that is sufficiently sensitive for diagnostic detection of DPV in acute infections, especially in tissues of ducks dying of the disease.

Modification of the passive heamagglutination test for detection of <i>infectious bursal disease virus</i>

“…Deng et al [14] described a Reverse Passive Haemagglutination (RPHA) test for detection of Duck plaque virus, another virus that lacks the haemagglutinin antigen. In RPHA, known specific antibody to the suspected virus is coupled to RBCs which are then added to dilutions of the suspected viral sample.…”

Duck virus enteritis (duck plague) – a comprehensive update

“…Pathogenesis of acute DEV infection has been studied in detail (Jansen 1968;Proctor, 1975Proctor, , 1976Deng et al 1984;Islam & Khan 1995;Hansen et al 2000;Shawky 2000;Shawky & Schat 2002;Sandhu & Shawky 2003;Cheng et al 2004Cheng et al , 2005Yuan et al 2005;Guiping et al 2007). Cheng et al (2004) demonstrated pathogenesis and immune mechanism of virulent and attenuated DEV, respectively.…”

“…The common immunological tests employed are neutralization assays, FAT and reverse passive hemagglutination test (RPHA) (Deng et al 1984;Sandhu & Shawky 2003). Next to virus inoculation in susceptible ducklings, FAT for the detection of virus on DEF and MDEF is considered most sensitive.…”

“…Deng et al (1984) developed an RPHA test to diagnose DEV. RPHA test has been found to be a rapid and easy procedure with adequate sensitivity in the tissues of ducks dying from acute infections, major drawback includes less sensitivity than either the plaque assays (PA) or the IF test.…”

The pathogenesis of duck virus enteritis in experimentally infected ducks: a quantitative time-course study using TaqMan polymerase chain reaction

“…The epithelial cells and lymphocytes of these organs are the principal sites of virus replication (Shawky, 2000;Yuan et al, 2005). Many studies on exploring the pathogenesis of acute DEV infection have been reported (Proctor, 1976;Deng et al, 1984;Lin et al, 1984;Plummer et al, 1998;Hansen et al, 2000;Cheng et al, 2004Cheng et al, , 2005. However, a great deal remains to be learned about quantitative aspects of DEV DNA load in various tissues and its relationship with the outcome of the infection.…”