Figure 1. Schematic representation of the herpes virion. A linear, double-strand deoxyribonucleic acid (DNA) molecule (red and orange) is surrounded by an icosahedral capsid (light blue), which in turn is covered by tegument (white). The outer surface of the tegument (grey) is associated with a lipid bilayer envelope (red and grey), which contains integral glycoproteins. Glycoprotein B trimer (green), glycoprotein C monomer (brown), glycoprotein D homodimer (light blue), glycoprotein H (purple) and glycoprotein L (orange) heterodimer allow the entry of the virion into the host cell.

Figure 1. Schematic representation of the herpes virion. A linear, double-strand deoxyribonucleic acid (DNA) molecule (red and orange) is surrounded by an icosahedral capsid (light blue), which in turn is covered by tegument (white). The outer surface of the tegument (grey) is associated with a lipid bilayer envelope (red and grey), which contains integral glycoproteins. Glycoprotein B trimer (green), glycoprotein C monomer (brown), glycoprotein D homodimer (light blue), glycoprotein H (purple) and glycoprotein L (orange) heterodimer allow the entry of the virion into the host cell.

Figure 2. Classes of viruses belonging to the family Hespesviridae based on the arrangement of repeated sequences in the genome. Human herpesvirus-6A and Human herpesvirus-6B (subfamily β-herpesvirinae) and Equine herpesvirus-2 (subfamily γ-herpesvirinae) genomes belong to class 1, while many members of the subfamily γ-herpesvirinae belong to class 2; members of the genus Varicellavirus (subfamily α-herpesvirinae) belong to class 3 genome, whereas Herpes simplex virus (subfamily α-herpesvirinae) and Cytomegalovirus (subfamily α-herpesvirinae) belong to class 4. The pol gene, which contains three conserved regions (A, B and C), is present in the architecture of all genome classes (illustrations not at scale). These regions encode highly conserved amino acid domains (DFA, ILK, TGV, IYG and KG1). PCR primers (dashed arrows) according to VanDevanter et al. are shown under the different class architectures. Primer’s directions are specified by dashed arrows. The product obtained by the Panherpes nested PCR is between region B and region C. The nomenclature used in all classes is U (unique), UL (unique long), US (unique short), TR (terminal repeat), TRL (terminal long repeat), IRL (internal long repeat), TRS (terminal short repeat), IRS (internal short repeat); TRL and TRS include a (terminal direct repeat) and IRL and IRS aI (internal inverse repeat). The orientations of repeated sequences are specified by block arrows.

Figure 2. Classes of viruses belonging to the family Hespesviridae based on the arrangement of repeated sequences in the genome. Human herpesvirus-6A and Human herpesvirus-6B (subfamily β-herpesvirinae) and Equine herpesvirus-2 (subfamily γ-herpesvirinae) genomes belong to class 1, while many members of the subfamily γ-herpesvirinae belong to class 2; members of the genus Varicellavirus (subfamily α-herpesvirinae) belong to class 3 genome, whereas Herpes simplex virus (subfamily α-herpesvirinae) and Cytomegalovirus (subfamily α-herpesvirinae) belong to class 4. The pol gene, which contains three conserved regions (A, B and C), is present in the architecture of all genome classes (illustrations not at scale). These regions encode highly conserved amino acid domains (DFA, ILK, TGV, IYG and KG1). PCR primers (dashed arrows) according to VanDevanter et al. are shown under the different class architectures. Primer’s directions are specified by dashed arrows. The product obtained by the Panherpes nested PCR is between region B and region C. The nomenclature used in all classes is U (unique), UL (unique long), US (unique short), TR (terminal repeat), TRL (terminal long repeat), IRL (internal long repeat), TRS (terminal short repeat), IRS (internal short repeat); TRL and TRS include a (terminal direct repeat) and IRL and IRS aI (internal inverse repeat). The orientations of repeated sequences are specified by block arrows.

Figure 3. Representation of the deoxyinosine-substituted primers generated by Ehlers et al. modifying the original degenerate primers and introducing deoxyinosine in all primer positions with 3- and 4-fold degeneration. For example, in the DFA degenerate primer, the degenerate N base was substituted with Ia.

Figure 3. Representation of the deoxyinosine-substituted primers generated by Ehlers et al. modifying the original degenerate primers and introducing deoxyinosine in all primer positions with 3- and 4-fold degeneration. For example, in the DFA degenerate primer, the degenerate N base was substituted with Ia.

Table 1. Macaviruses (shaded) and non Macavirus Malignant Catarrhal Fever Viruses, their clinical significance, reservoir(s) and susceptible species. Based on the ICTV classification at time of printing.

Table 1. Macaviruses (shaded) and non Macavirus Malignant Catarrhal Fever Viruses, their clinical significance, reservoir(s) and susceptible species. Based on the ICTV classification at time of printing.

VirusClinical MCFReservoir
SpeciesSusceptible SpeciesReferenceAlcelaphine γ- herpesvirus-1YesWildebeestCattle
Deer[14]Alcelaphine γ- herpesvirus-2YesHartebeest
TopiBarbary red deer
Bison[14]Ovine γ-herpesvirus-2YesSheep
MouflonCattle
Water buffalo
Moose
Domestic goats
Deer
Bison
Pig
Giraffe[1,14]Caprine γ-herpesvirus-2YesGoatSika deer
White-tailed deer[14]Hippotragine γ-herpesvirus-1NoRoan antelopeNo susceptible species identified[14]Bovine γ-herpesvirus-6NoCattleNo susceptible species identified[17]Suid γ-herpesvirus-3NoPigNo susceptible species identified[18]Suid γ-herpesvirus-4NoPigNo susceptible species identified[18]Suid γ-herpesvirus-5NoPigNo susceptible species identified[18]MCFV–white-tailed deer/Caprine
γ-herpesvirus-3YesGoatWhite-tailed deer
Red brocket deer
Reindeer[14]Ibex-MCFVYesIbexBongo antelopes
Anoa
Pronghorn
Duikers[14,19]

Table 2. Primers sequences employed for the Degenerate/Consensus PCR according VanDevanter et al. (2a) and the Consensus Panherpes PCR according to Ehlers et al. (2b). Primers used for Sanger Sequencing following the Degenerate/Consensus PCR according to VanDevanter et al. are shown in italic. Degenerate and inosine-substituted equivalents are shown in bold. Ia = Deoxyinosine.

Table 2. Primers sequences employed for the Degenerate/Consensus PCR according VanDevanter et al. (2a) and the Consensus Panherpes PCR according to Ehlers et al. (2b). Primers used for Sanger Sequencing following the Degenerate/Consensus PCR according to VanDevanter et al. are shown in italic. Degenerate and inosine-substituted equivalents are shown in bold. Ia = Deoxyinosine.

2a. Consensus Panherpes Polymerase Chain Reaction PrimersFirst PCRForwardReverseDFA: 5′-GAY TTY GCN AGY YTN TAY CC-3′KG1: 5′-GTC TTG CTC ACC AGN TCN ACN CCY TT-3′ILK: 5′-TCC TGG ACA AGC AGC ARN YSG CNM TNA A-3I’Second PCRForwardReverseTGV: 5′-TGT AAC TCG GTG TAY GGN TTY ACN GGN GT-3′ IYG: 5′-CAC AGA GTC CGT RTC NCC RTA DAT-3′. Approximate product: 215- to 315-bpSanger SequencingForwardReverseTGVseq: 5′-CAT CTG ATG TAA CTC GGT GTA-3′ bottom lineIYGseq: 5′-GAC AAA CAC AGA GTC CGT-3′2b. Consensus Panherpes PCR with Consensus/Degenerate
and Deoxyinosine-Substituted PrimersFirst PCRForwardReverseDFA: 5′-GAY TTY GCN AGY YTN TAY CC-3′KG1: 5′-GTC TTG CTC ACC AGN TCN ACN CCY TT-3′ILK: 5′-TCC TGG ACA AGC AGC ARN YSG CNM TNA A-3′Deoxyinosine-substituted equivalentDeoxyinosine-substituted equivalentDFA: 5′-GAY TTY GCIa AGY YTIa TAY CC-3I′KG1: 5′-GTC TTG CTC ACC AGIa TCIa ACIa CCY TT-3′ILK: 5′-TCC TGG ACA AGC AGC ARIa YSG CIaM TIaA-3′Second PCRForwardReverseTGV: 5′-TGT AAC TCG GTG TAY GGN TTY ACN GGN GT-3′IYG: 5′-CAC AGA GTC CGT RTC NCC RTA DAT-3′Deoxyinosine-substituted equivalent
TGV: 5′-TGT AAC TCG GTG TAY GGIa TTY ACIa GGIa GT-3′Deoxyinosine-substituted equivalent
IYG: 5′-CAC AGA GTC CGT RTC IaCC RTA IaAT-3′

Table 3. Primer sequences for the Semi nested consensus PCR for the amplification of fragment of the glycoprotein B according to Chmielewicz et al. (3a) and Degenerate/Consensus primers and Deoxyinosine-substituted primers used by Ehlers et al. in the gB nested PCR (3b). Ia = Deoxyinosine.

Table 3. Primer sequences for the Semi nested consensus PCR for the amplification of fragment of the glycoprotein B according to Chmielewicz et al. (3a) and Degenerate/Consensus primers and Deoxyinosine-substituted primers used by Ehlers et al. in the gB nested PCR (3b). Ia = Deoxyinosine.

3a. Semi-Nested PCR for the Glycoprotein B (gB) GeneFirst PCRForwardReverse702 Gb: 5′-CAR IaTIa CAR TWT GCM TAY GAC-3′702 Gb: 5′-GTA RTA RTT RTA YTC YCT RAA-3′Second PCRForwardReverse734 gB: 5′-GCA AAA TCA ACC CTA CVA GYG TNA TG-3′702 gB: 5′-GTA RTA RTT RTA YTC YCT RAA-3′Approximate product: 515bp3b. Nested PCR for the Glycoprotein B (gB) GeneFirst PCRForwardReverseGH1 2759:
5′-CCT CCC AGG TTC ART WYG CMT AYG A-3IGH1 2762:
5′-CCG TTG AGG TTC TGA GTG TAR TAR TTR TAY TC-3′GH2 3029:
5′-CCC AGT TGC ART WYG GC(N/Ia) TAY GA-3′GH2 3033:
5′-GCC AGG CGT TGC GT(N/Ia) TAR TAR TTR TA-3′Second PCRForwardReverseGH1 2760:
5′-AAG ATC AAC CCC AC(N/Ia) AG(N/Ia) GT(N/Ia) ATG-3′GH1 2761:
5′-GTG TAG TAG TTG TAC TCC CTR AAC AT(N/Ia) GTY TC-3′ GH2 3031:
5′-CAA GAT TAA CCC CAC (N/Ia)AG (N/Ia)GT (N/Ia)AT G-3′GH2 3032:
5′-TTG CGT GTA GTA GTT GTA YTC (N/Ia)CT RAA CAT-3′