Up to now, the earliest proof of PCV2 in swine was reported from a German sample from 1962 [
5]. In this specimen, PCV2 detection was positive using DNA in situ hybridization and PCR. However, since only a 66 bp sequence of ORF1 (GenBank EU158775) is available in this case, the genotype cannot be specified. One can speculate whether this is the first evidence of PCV2a, as it is believed that this genotype is the first to circulate in the pig population worldwide and has been linked to the onset of the postweaning multisystemic wasting syndrome (PMWS) [
6‐
9]. In the first period, PCV2a was the most prevalent PCV2 genotype in clinically affected pigs. This changed with the rise of a second cluster of PCV2 variants, eventually called PCV2b [
10]. Initially in Europe, later in North America, an increased incidence of PCV2 associated diseases (PCVD) was observed in the early 2000s, this time caused by the genotype PCV2b, which replaced PCV2a as the dominant virus variant in PCVD outbreaks [
8,
11‐
13]. This change is commonly known as “PCV2 genotype shift”. For several years PCV2b dominated the world as causative agent of PCVD [
3,
14‐
16] and caused severe economic losses. In 2008, Dupont and colleagues published a retrospective study of PCV2 isolates from Danish archives. Within this collection, three genome sequences obtained from samples collected in 1980, 1987 and 1990 did not cluster with known PCV2a or 2b strains, but were assigned to a new branch of the PCV2 phylogenetic tree, later referred to as genotype PCV2c [
8]. Interestingly, after their discovery, PCV2c strains were not detected in swine populations worldwide for a long period. More recently, PCV2c was detected in feral pigs in the Brazilian Pantanal [
17] and a Chinese group reported potential recombinants between PCV2b and PCV2c strains [
18]. However, for global swine industry PCV2c is still of minor interest today. In contrast, genotype PCV2d is a challenge and a serious economic problem in pig production systems all over the world. It has become the most prevalent PCV2 genotype (“second genotype shift”), at least in clinically affected animals. About a decade ago, Guo and colleagues reported an emerging “mutant” PCV2 (strain BDH, HM038017) characterized by a mutation in the stop codon of ORF2 resulting in a capsid protein of 234 amino acid (aa) and which showed higher levels of virulence compared to classical PCV2a or 2b strains [
19,
20]. Remarkably, other groups could not confirm a particular virulence of this virus variant [
21]. However, BDH-like viruses spread quickly to North America [
22], Europe [
23] and the rest of the world [
24]. In the phylogenetic tree of PCV2, these viruses formed a separate cluster together with virus strains originally described by Olvera and colleagues as subgroup 1C (= PCV2b, subgroup 1C) [
13]. Therefore, different names were used for this group for some time (mutant PCV2, mPCV2b, PCV2b-1C), but eventually this cluster was accepted as the genotype PCV2d [
2]. Although the PCV2d reference strain BDH was isolated in 2008 [
19], the oldest published PCV2d sequence (AY484410) was obtained from a Dutch study carried out in 2001/2002 [
25]. Furthermore, Xiao and colleagues demonstrated a significant heterogeneity within the genotype PCV2d and subdivided it into the major clades PCV2d-1 (containing mainly “old” sequences, obtained before 2008) and PCV2d-2 (BDH-like strains) [
24]. This classification scheme was used in several other studies [
2,
26,
27] until the new genotyping methodology [
4] transferred most of the PCV2d-1 strains into the new genotype PCV2g. Interestingly, some of the former PCV2d-1 members (e.g. AY484410 or AY181946) remained within in the “new” genotype 2d. Remarkably, most PCV2g strains have been isolated from wild boars. In 2016, Davies and colleagues reported a unique new cluster of PCV2 strains, collected in North America from 2006 to 2015. These viruses were characterized by a larger genome size due to an insertion at the 3′ end of ORF2, causing a 238 aa capsid protein (see Table
1). This cluster, named genotype PCV2e, showed the highest distance to all other PCV2 genotypes, and it was hypothesized that it might be a kind of PCV2 ancestor [
28]. Meanwhile, PCV2e strains have also been detected in China [
29] and Japan (GenBank number LC278353). Bao and colleagues introduced genotype PCV2f in 2018, containing mainly strains from China, India and Indonesia [
30]. Two isolates that are an exception were reported from a wild boar in Croatia (HQ591381) and feral pigs in Brazil (KJ094600), originally described as PCV2a [
17]. The latter is showing a comparative big distance to the other PCV2f members. As described above, genotype PCV2g includes strains previously described as PCV2d-1, having a capsid protein of 234 aa. However, the analysis of Franzo and Segales [
4] shows that this group also contains PCV2 sequences with a 233 aa ORF2 product and a total length of 1768 bp, which is typical for PCV2a strains. This might explain, why some of these strains were described as “inter-genotype” recombinants before [
31]. One of the oldest members of this new genotype, sequence AY713470, is a good example of how complex correct genotyping of PCV2 strains can be. This strain originated from a German wild boar, which was hunted in the winter of 2003/04. The longer capsid protein (234 aa) and the full genome length of 1767 bp of the virus were explained by a one nucleotide deletion at the end of ORF2 of a PCV2a virus [
32]. In 2007, Olvera and colleagues classified this virus as a member of group 1C [
13], and after the proposal of Segales and colleagues for PCV2 genotype definition and nomenclature [
10] it was reclassified as PCV2b. Three years later it was integrated into a putative new genotype PCV2d [
19], but alternative names (e.g. PCV2b-1C, mPCV2b) were still used [
22,
23,
33] until the revision of PCV2 genotyping by Franzo and colleagues in 2015, which exemplified the classification to genotype PCV2d [
2]. In the same year Xiao and colleagues introduced the virus as one of the PCV2d-1 reference strains [
24], until recently it became one of the reference strains for genotype PCV2g [
4]. Simultaneously with PCV2g, the new genotype PCV2h was documented. Up to now, only PCV2 sequences from Asia belong to this group [
4]. Some of these strains were previously described as recombinant viruses or as “intermediate” strains [
24,
34,
35].
From the beginning, scientists around the world tried to correlate different phylogenetic clusters or genotypes of PCV2 with characteristics such as virulence or clinical manifestation [
8,
13,
36]. Although virulence-contributing features of the PCV2 genome or proteome are still poorly understood, correct PCV2 genotyping has become an important task, not only for answering scientific questions, but also in routine diagnostic work. Molecular epidemiology, the detection of emerging genotypes, tracing PCV2 spreading by international and national trade, the investigation of viral evolution and the evaluation of vaccine effectiveness and cross protection are only a few points, which illustrate the importance of correct PCV2 genotyping [
2,
3,
37,
38]. Because of its small, circular genome, full genome sequencing of PCV2 is comparatively easy. Several protocols to amplify and sequence overlapping genome fragments have been published [
8,
12,
25,
39]. In general, samples from animals with PCVD can be targeted this way because of the high viral load in these specimens. Receiving high quality sequencing results from subclinically infected pigs is more challenging. Several attempts to increase sensitivity by nested PCR or unspecific amplification of the circular DNA of circoviruses by rolling circling amplification (RCA) before PCR have been published [
14,
40]. However, real-time PCR is a valuable option for faster, cheaper, more sensitive and more mass compatible genotyping in routine diagnostics. Two multiplex TaqMan-based qPCR assays for the differentiation of PCV2a and PCV2b were published already a decade ago [
41,
42]. With the rise of PCV2d as new dominant genotype, these assays lost their importance, especially, as in one of the assays, cross reaction of the PCV2b specific TaqMan probe with PCV2d strains was observed [
22].