Comparison Of Four Anti-Avian IgY Secondary Antibodies Used In Western Blot And Dot-Blot ELISA To Detect Avian Bornavirus Antibodies In Four Different Bird Species

Figures & data

Figure 1 Representative membranes of Western blot results using anti-macaw, anti-bird, anti-duck, and anti-chicken IgY secondary antibody on ABV antibody positive and negative plasma from Blue and gold macaw (Ara ararauna), Cockatiel (Nymphicus hollandicus), and Mallard (Anas platyrhynchos).

Notes: The N-protein signal was located at approximately 38–41 kDa.
Abbreviations: 2° Ab, secondary antibody; MW, molecular weight.

Figure 2 Representative membranes of dot-blot ELISA results using anti-macaw, anti-bird, anti-duck, and anti-chicken IgY secondary antibody on ABV antibody positive and negative plasma from Blue and gold macaw (Ara ararauna), Cockatiel (Nymphicus hollandicus), Monk parakeet (Myiopsitta monachus), and Mallard (Anas platyrhynchos).

Abbreviation: 2° Ab, secondary antibody

Figure 3 Digital image analysis of dot-blot ELISA used to generate the signal intensity value.

Notes: The dot-blot ELISA with the delineated area of interest highlighted in the red rectangle, the graphic display, and the signal intensity value are displayed using samples from (A) Blue and gold macaw (Ara ararauna), (B) Cockatiel (Nymphicus hollandicus), (C) Monk parakeet (Myiopsitta monachus), and (D) Mallard (Anas platyrhynchos).

Table 1 Signal Intensities Obtained When Testing ABV Antibody Positive And Negative Plasma From Blue And Gold Macaw (Ara ararauna), Cockatiel (Nymphicus hollandicus), Monk Parakeet (Myiopsitta monachus), And Mallard (Anas platyrhynchos) With A Dot-Blot ELISA Using Anti-Macaw, Anti-Bird, Anti-Duck, And Anti-Chicken IgY Secondary Antibody

Bird Species	2° Antibody	N	Signal Intensity
			Positive Sample	Negative Sample
Blue and gold macaw
	Anti-macaw	6	19,461^a (±1,338)	318* (±135)
	Anti-bird	6	2,549^b (±462)	1,080 (±786)
	Anti-duck	6	92^c (±40)	148 (±76)
	Anti-chicken	6	415^c (±148)	152 (±52)
Cockatiel
	Anti-macaw	6	21,032^a (±1218)	221* (±116)
	Anti-bird	6	3,690^b (±459)	109* (±18)
	Anti-duck	6	180^c (±54)	127 (±63)
	Anti-chicken	6	435^c (±251)	94 (±43)
Monk parakeet
	Anti-macaw	6	17,785^a (±830)	805* (±176)
	Anti-bird	6	15,522^b (±1032)	703* (±136)
	Anti-duck	6	1,389^c (±355)	739 (±267)
	Anti-chicken	6	736^c (±88)	1,328 (±381)
Mallard
	Anti-macaw	6	561^a (±333)	297 (±87)
	Anti-bird	6	12,724^b (±2459)	814* (±514)
	Anti-duck	6	18,098^c (±2102)	146* (±51)
	Anti-chicken	6	5,751^d (±714)	846* (±393)

Notes: Signal intensity data are presented as the mean (±SEM). ^a,b,c,d Different superscript letters within each bird species sampled indicate a significant difference (P<0.05) between secondary antibody values. *Negative sample value is significantly different (P<0.05) from the corresponding positive sample value of the same secondary antibody. The negative values within each bird species sampled were not significantly different.

Abbreviations: ABV, avian bornavirus; 2° antibody, secondary antibody; N, sample size.