Table 3.

A summary of all methods used in screening for BFDV in wild and captive psittacine populations, a count of how many published studies in which each has been used and example publications for where each has been applied

Method	Description	Times used	Example references
Blocking ELISA	A blocking ELISA is a method used to immobilize biomolecules, primarily proteins, to a plate via passive or covalent interactions, minimising nonspecific binding to the surface by saturating unoccupied binding sites with a blocking reagent	1	[109]
DNA in situ hybridization	DNA in situ hybridization is a technique used in the localisation of specific nucleic acid targets within fixed tissues and cells using an oligonucleotide probe before microscopically visualizing the results	4	[69, 81]
Dot-blot DNA hybridization	Dot blot hybridization is a technique used to determine the abundance of certain DNA in an extraction dotted onto a membrane through hybridization with universal and specific oligonucleotide probes	2	[20, 21]
Duplex shuttle PCR	Duplex shuttle PCR is a process that allows the co-amplification of separate regions of a gene in a single PCR reaction using different pairs of primers in the same reaction mixture	1	[51]
Endocrinological response	Endocrinological response is a method used to challenge the host immune system with a hormone that encourages the production and release of a stress hormone to evaluate whether any differences exist between healthy and infected individuals	1	[87]
Haemagglutination assay	Haemagglutination assay (HA) is a method used to quantify the amount of virus attached to molecules on the surface of host red blood cells in a series of dilutions of a viral suspension	12	[26, 113]
Haemagglutination inhibition	A modified version of the HA where a standard amount of virus and host blood cells are used with the addition of a serially diluted antiserum to determine which concentration inhibits agglutination of the cells	12	[9, 65]
Haematology	Haematology is the study of the morphology and physiology of blood and, in this context, relates to the diagnosis and monitoring of pathogens present in the blood stream	3	[87, 89]
Histology	Histology is the microscopic examination of stained tissues and is applied in the screening for BFDV to determine if viral inclusion bodies are present. Techniques include light and electron microscopy	28	[57, 114]
Immunohistochemical tests	Immunohistochemistry (IHC) is a technique used to observe the physical characteristics of antibodies and their concentration and distribution within host tissue. In screening for BFDV, specimens are stained using the avidin-biotin complex (ABC) immunoperoxidase technique and then exposed to a primary antibody	5	[19, 91]
Quantitative (real-time) PCR	Quantitative (or real-time) polymerase chain reaction (qPCR) is a technique used to both amplify and quantify target DNA through the use of either nonspecific fluorescent dyes that intercalate with double-stranded DNA or a sequence-specific fluorescent probe that hybridizes with the target	6	[37, 54]
Standard PCR	Polymerase chain reaction (PCR) is a technology used to amplify a piece of DNA across several orders of magnitude through a process of thermal cycling in combination with oligonucleotide probes synthesised to bind to the target region and a DNA polymerase enzyme	41	[48, 102]
Virus purification	Virus purification allows the careful study of viral synthesis within cells by combining ultracentrifugation, adsorption chromatography, electrophoresis, and partition in liquid phases to separate virions from incomplete protein fragments and cell debris	3	[26, 75]
Whole-genome sequencing	Whole-genome sequencing is a laboratory process that determines the complete DNA sequence of an organism’s genome at a single time and can be used for multiple tissue types and when only very small quantities of a full DNA copy are present	23	[115, 116]