Table 2.

Methods for Maize dwarf mosaic virus (MDMV) detection.

Method	Platform	Effect on MDMV Variation	Reliability	Efficiency	References
Indirect Enzyme-Linked Immunosorbent Assay (ELISA)	Rabbit anti-mouse IgM capture monoclonal antibody Rabbit anti-mouse IgG second monoclonal antibody	MDMV-A and MDMV-B	Allow detection of MDMV-A and MDMV-B bound IgG among tested leaves sap infected with several different strains of MDMV and Sugarcane mosaic virus (SCMV).	MDMV-A-specific ELISA = antigen detected in sap at a dilution of 1:60 with optimal sample pH (7.5–8.5). MDMV-B-specific ELISA = detect antigen in sap at dilution end point of 1:2560	[91]
Indirect Enzyme-Linked Immunosorbent Assay (ELISA)	M-C antiserum M-D antiserum	M-C and M-D	M-C antigen reacts strongly with M-A, M-D, Sorghum red stripe virus (SRV) antisera. M-D antigen reacts strongly with M-D, M-C antisera.	M-C particles react with M-A and SRV antisera in the range of 1:128 to 1:512 dilution end points, with M-D antisera at dilution end point of 1:4. M-D antigen reacted with homologous antisera, M-D up to a dilution of 1:512, with M-C antisera at 1:16 dilution.	[88]
Double-Antibody Sandwich Enzyme-Linked Immunosorbent Assay (DAS ELISA)	Anti-MDMV-A rabbit serum purified for IgG Alkaline phosphatase coupled to purified IgG as conjugate	MDMV-A	Visual evaluation at absorbance of 405 nm gives reliable information on MDMV-A presence in leaves extracts tested.	Visible yellow colour formed in wells with antigen diluted up to 10−4. Sensitivity is 100 times better than conventional infectivity test.	[92]
Double-Antibody Sandwich Enzyme-Linked Immunosorbent Assay (DAS ELISA)	IgG purified from polyclonal anti-MDMV serum Alkaline phosphatase coupled to purified IgG-E as conjugate	MDMV-A, MVMV-J, MDMV-L, MDMV-SP, MDMV-YU	Among all reference strains, only MDMV strains react positively in DAS-ELISA (recorded OD 405 nm values at least twice the healthy sap OD 405 nm values) with anti-MDMV IgG.	IgG antibody dilution to 1 μg/mL able to detect MDMV antigen dilution to 1/100.	[30]
Double-Antibody Sandwich Enzyme-Linked Immunosorbent Assay (DAS ELISA)	MDMV-Arg (Argentina strain) [93] specific polyclonal IgG Alkaline phosphatase-conjugated IgG as secondary antibody	MDMV	DAS-ELISA absorbance values of infected maize leaf samples from field grown plants and maize leaf sample with MDMV-Arg isolate were highly significant while the values from healthy control and buffer were very low.	MDMV-Arg specific polyclonal IgG strongly reactive up to 1:2000 dilution with MDMV antigen from infected field, Chile.	[19]
Capture Reverse Transcription-Polymerase Chain Reaction (RT-PCR)	Primers designed to MDMV-Arg [93] capsid protein gene	MDMV	Single band of expected size (1104 bp) obtained.	Detect MDMV samples from Chile which closely related to MDMV-Arg (Argentina strain) efficiently.	[19]
Reverse Transcription-Polymerase Chain Reaction (RT-PCR)	Fwd primer = oligo1n: ATGGTHTGGTGYATHGARAAYGG Rvs primer = oligo2n: TGCTGCKGCYTTCATYTG *single lettercode: H = A/C/T, Y = C/T R = A/G, K = G/T	MDMV-SP, MDMV-Bu, MDMVJIL	Single product of expected size (327 nts) obtained for all tested MDMV isolates.	Efficient for dealing with well-characterized strains, field collected isolates.	[30]
Reverse Transcription-Polymerase Chain Reaction (RT-PCR)	Universal primer (Sprimer: 5′-GGXAAYAAYAGYGGXCAZCC-3′, X = A/G/C/T; Y = T/C; Z = A/G) M4 primer	MDMV	cDNA fragments of expected size were amplified from the 3′ terminus of RNA genomes of 21 different viruses under family potyviridae including MDMV.	Universal primer designed based on an alignment of the amino acid sequences around the conserved GNNSGQP motif in nuclear inclusion body b (NIb) gene of family potyviridae members. Hence, it is proved useful for detection of potyviridae members.	[94]
Reverse Transcription-Polymerase Chain Reaction (RT-PCR)	MDMV F1: 5′-CAACCAGGGCYGAATTTGATAG-3′ MDMV R1: 5′-GTGCAAGGC TRAAGTCGG TTA-3′	MDMV	Supposed to yield a PCR product of expected size (336 bp)	MDMV can be distinguishable from Sugarcane mosaic virus, SCMV and Johnsongrass mosaic virus, JGMV through RT-PCR	[57]
Combined Reverse Transcription Polymerase Chain Reaction (RT-PCR) with Electrochemiluminescence method	Specific nucleic acid sequences (20 bp) were added to 5′ terminal of all primers Biotin was introduced into reverse primer	MDMV	PCR yielded a product with a single band of expected size (643 bp) for all 4 tested viruses including MDMV.	This method has higher sensitivity and lower cost than others. It can effectively detect the MDMV with simplicity and stability.	[95]
Competitive Radioimmunoassay (RIA)	Rabbit anti-mouse monoclonal IgG	MDMV-A (referred here as MDMV-AP) and MDMV-B	Feasible alternative to the use of polyclonal antisera in detecting homologous viruses (MDMV, Soybean mosaic, SMV, Lettuce mosaic virus, LMV).	Antigen (purified virus) detected at dilution of 10–50 ng/mL.	[96]
Dot Blot Immunoassay	MDMV-Arg [93] polyclonal antiserum	MDMV	MDMV symptomatic field grown plants had strong reaction with the polyclonal antiserum against MDMV-Arg isolate while healthy plants were negative.	MDMV samples reacted with MDMV-Arg polyclonal antiserum of dilution up to 1:5000	[19]
Sodium dodecyl sulfate (SDS) immunodiffusion test	M-D antiserum M-D antiserum	MDMV-C and MDMV-D	M-C antiserum reacted with both M-C and M-D antigens forming a spur, which indicate partial serological relatedness. M-D antiserum reacted with its homologous viral antigen, M-D.	No precipitin lines were obtained when antisera reacted against healthy crude sap	[88]
DNA Microarray (Maizepath)-based detection	Microarray with 60-mer oligonucleotide probes complementary to genomes of 5 maize pathogens including MDMV	MDMV	Obtained results indicate that the fluorescence signals from MDMV, other pathogens and control probes are well distinguished in all performed experiments.	Gives more than 180 K probes in total, thereby classified as high-density microarray that able to investigate thousands of genomic loci in a high-resolution manner.	[97]