Fassbinder-Orth Carol A, Wilcoxen Travis E, Tran Tiffany, Boughton Raoul K, Fair Jeanne M, Hofmeister Erik K, Grindstaff Jennifer L, Owen Jen C
Biology Department, Creighton University, 2500 California Plaza, Omaha, NE 68178, USA.
Biology Department, Millikin University, 1184 West Main Street, Decatur, IL 62522, USA.
Methods Ecol Evol. 2016 Oct;7(10):1174-1181. doi: 10.1111/2041-210X.12583. Epub 2016 May 28.
Immunological reagents for wild, non-model species are limited or often non-existent for many species.In this study, we compare the reactivity of a new anti-passerine IgY secondary antibody with existing secondary antibodies developed for use with birds. Samples from 41 species from the following six avian orders were analysed: Anseriformes (1 family, 1 species), Columbiformes (1 family, 2 species), Galliformes (1 family, 1 species), Passeriformes (16 families, 34 species), Piciformes (1 family, 2 species) and Suliformes (1 family, 1 species). Direct ELISAs were performed to detect total IgY using goat anti-passerine IgY, goat anti-chicken IgY or goat anti-bird IgY secondary antibodies.The anti-passerine antibody exhibited significantly higher IgY reactivity compared to the anti-chicken and/or anti-bird antibodies in 80% of the passerine families tested. Birds in the order Piciformes (woodpeckers) and order Suliformes (cormorants) were poorly detected by all three secondary antibodies. A comparison of serum and plasma IgY levels was made within the same individuals for two passerine species (house finch and white-crowned sparrow), and serum exhibited significantly more IgY than the plasma for all three secondary antibodies. This result indicates that serum may be preferred to plasma when measuring total antibody levels in blood.This study indicates that the anti-passerine IgY secondary antibody can effectively be used in immunological assays to detect passerine IgY for species in most passerine families and is preferred over anti-chicken and anti-bird secondary antibodies for the majority of passerine species. This anti-passerine antibody will allow for more accurate detection and quantification of IgY in more wild bird species than was possible with previously available secondary antibodies.
对于许多野生非模式物种而言,免疫试剂十分有限,甚至常常不存在。在本研究中,我们比较了一种新型抗雀形目IgY二抗与现有的用于鸟类的二抗之间的反应性。分析了来自以下六个鸟类目41个物种的样本:雁形目(1科,1种)、鸽形目(1科,2种)、鸡形目(1科,1种)、雀形目(16科,34种)、啄木鸟目(1科,2种)和鹈形目(1科,1种)。使用山羊抗雀形目IgY、山羊抗鸡IgY或山羊抗鸟类IgY二抗进行直接ELISA检测总IgY。在80%检测的雀形目科中,抗雀形目抗体显示出比抗鸡和/或抗鸟类抗体显著更高的IgY反应性。啄木鸟目(啄木鸟)和鹈形目(鸬鹚)的鸟类被所有三种二抗检测的效果都很差。对两种雀形目物种(家朱雀和白冠雀)的同一个体的血清和血浆IgY水平进行了比较,对于所有三种二抗,血清中的IgY都显著多于血浆。该结果表明,在测量血液中的总抗体水平时,血清可能比血浆更受青睐。本研究表明,抗雀形目IgY二抗可有效地用于免疫测定,以检测大多数雀形目科物种的雀形目IgY,并且对于大多数雀形目物种而言,它比抗鸡和抗鸟类二抗更受青睐。与以前可用的二抗相比,这种抗雀形目抗体将能够更准确地检测和定量更多野生鸟类物种中的IgY。