Background
Methods
Specimen collection
DNA extraction and parvovirus screening
SimpleProbe® assay for CPV-2 genotyping
VP2 gene amplification, sequencing and sequence analyses
Results
SYBR-based real-time PCR amplification
Genotype analysis and geographical distribution of CPV-2 variants
CPV-2 genotype | SimpleProbe® analysis | Reference melting temperature (CI95%) [26] | |
---|---|---|---|
Number | Melting temperature of Vietnamese samples (CI95%) | ||
2a | 6 | 50.23 (50.2–50.3) | 50.2 (50.1–50.5) |
2b | 0 | N/Aa | 57.8 (57.7–58.5) |
2c | 251 | 52.75 (52.7–52.8) | 52.3 (52.2–53.2) |
Genotype of CPV-2 vaccinated dogs
Amino acid and nucleotide sequence analysis of VP2
Strains | Position of nucleotide sequence (amino acid) | |||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
13–15 (5) | 259–261 (87) | 799–801 (267) | 898–900 (300) | 964–966 (322) | 967–969 (323) | 970–972 (324) | 1000–1002 (334) | 1021–1023 (341) | 1108–1110 (370) | 1276–1278 (426) | 1318–1320 (440) | 1339–1341 (447) | 1663–1665 (555) | |
Reference strain | ||||||||||||||
CPV-2 | ||||||||||||||
M38245 (USA) | GCA (Ala) | ATG (Met) | TTT (Phe) | GCT (Ala) | ACA (Thr) | AAC (Asn) | TAT (Tyr) | GCT (Ala) | CCA (Pro) | CAA (Gln) | AAT (Asn) | ACA (Thr) | ATA (Ile) | GTA (Val) |
CPV-2a | ||||||||||||||
M24000 (USA) | GCA (Ala) | TTG (Leu) | TTT (Phe) | GGT (Gly) | ACA (Thr) | AAC (Asn) | TAT (Tyr) | GCT (Ala) | CCA (Pro) | CAA (Gln) | AAT (Asn) | ACA (Thr) | ATA (Ile) | ATA (Val) |
FJ197847 (vaccine) | GCA (Ala) | ATG (Met) | TTT (Phe) | GGT (Gly) | ACA (Thr) | AAC (Asn) | TAT (Tyr) | GCT (Ala) | CCA (Pro) | CAA (Gln) | AAT (Asn) | ACA (Thr) | ATA (Ile) | GTA (Val) |
New CPV-2a | ||||||||||||||
AY742953 (USA) | GCA (Ala) | TTG (Leu) | TTT(Phe) | GGT (Gly) | ACA (Thr) | AAC (Asn) | TAT (Tyr) | GCT (Ala) | CCA (Pro) | CAA (Gln) | AAT (Asn) | ACA (Thr) | ATA (Ile) | GTA (Val) |
KF676668 (China) | GCA (Ala) | TTG (Leu) | TAT(Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CAA (Gln) | AAT (Asn) | GCA (Ala) | ATA (Ile) | GTA (Val) |
KR611488 (China) | GCA (Ala) | TTG (Leu) | TAT(Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CAA (Gln) | AAT (Asn) | GCA (Ala) | ATA (Ile) | GTA (Val) |
JQ686671 (China) | GCA (Ala) | TTG (Leu) | TAT(Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CAA (Gln) | AAT (Asn) | GCA (Ala) | ATA (Ile) | GTA (Val) |
CPV-2b | ||||||||||||||
M78452 (USA) | GCA (Ala) | TTG (Leu) | TTT (Phe) | GGT (Gly) | ACA (Thr) | AAC (Asn) | TAT (Tyr) | GCT (Ala) | CCA (Pro) | CAA (Gln) | GAT (Asp) | ACA (Thr) | ATA (Ile) | GTA (Val) |
New CPV-2b | ||||||||||||||
AY742955 (USA) | GCA (Ala) | TTG (Leu) | TTT(Phe) | GGT (Gly) | ACA (Thr) | AAC (Asn) | TAT (Tyr) | GCT (Ala) | CCA (Pro) | CAA (Gln) | GAT (Asp) | ACA (Thr) | ATA (Ile) | GTA (Val) |
JQ268284 (China) | GCA (Ala) | TTG (Leu) | TAT(Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CAA (Gln) | GAT (Asp) | GCA (Ala) | ATA (Ile) | GTA (Val) |
KR611459 (China) | GCA (Ala) | TTG (Leu) | TAT(Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CAA (Gln) | GAT (Asp) | GCA (Ala) | ATA (Ile) | GTA (Val) |
KR611461 (China) | GCA (Ala) | TTG (Leu) | TAT(Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CAA (Gln) | GAT (Asp) | GCA (Ala) | ATA (Ile) | GTA (Val) |
CPV-2c | ||||||||||||||
FJ005235 (USA) | GCA (Ala) | TTG (Leu) | TTT (Phe) | GCT (Ala) | ACA (Thr) | AAC (Asn) | TAT (Tyr) | GCT (Ala) | CCA (Pro) | CAA (Gln) | GAA (Glu) | ACA (Thr) | ATA (Ile) | GTA (Val) |
FJ222821 (Italy) | GCA (Ala) | TTG (Leu) | TTT (Phe) | GGT (Gly) | ACA (Thr) | AAC (Asn) | TAT (Tyr) | GCT (Ala) | CCA (Pro) | CAA (Gln) | GAA (Glu) | ACA (Thr) | ATA (Ile) | GTA (Val) |
KY937650 (China) | GGA (Gly) | TTG (Leu) | TAT(Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CAA (Gln) | GAA (Glu) | ACA (Thr) | ATA (Ile) | GTA (Val) |
KU244254 (Taiwan) | GCA (Ala) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATA (Ile) | GTA (Val) |
AB120727 (Vietnam) | GCA (Ala) | TTG (Leu) | TTT (Phe) | GGT (Gly) | ACA (Thr) | AAC (Asn) | TAT (Tyr) | GCT (Ala) | CCG (Pro) | CAA (Gln) | GAA (Glu) | ACA (Thr) | ATA (Ile) | GTA (Val) |
This study | ||||||||||||||
CPV-2a | ||||||||||||||
DN33/MK357724 | GCA (Ala) | ATG (Met) | TTT (Phe) | GCT (Ala) | ACA (Thr) | AAC (Asn) | TAT (Tyr) | GCT (Ala) | CCA (Pro) | CAA (Gln) | AAT (Asn) | ACA (Thr) | ATA (Ile) | GTA (Val) |
CPV-2c | ||||||||||||||
HN7AA/MK357734 | GGA (Gly) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATA (Ile) | GTA (Val) |
HN4AA/MK357735 | GGA (Gly) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCC (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATG (Met) | GTA (Val) |
N11AA/MK357736 | GGA (Gly) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATG (Met) | GTA (Val) |
HN37AA/K357737 | GGA (Gly) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATG (Met) | GTA (Val) |
HCM4/MK357725 | GGA (Gly) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATG (Met) | GTA (Val) |
HCM16/MK357726 | GGA (Gly) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATG (Met) | GTA (Val) |
HCM22/MK357727 | GGA (Gly) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATA (Ile) | GTA (Val) |
HCM25/MK357728 | GGA (Gly) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATG (Met) | GTA (Val) |
HCM27/MK357729 | GGA (Gly) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATG (Met) | GTA (Val) |
HCM32/MK357730 | GGA (Gly) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATG (Met) | GTA (Val) |
HCM48/MK357731 | GGA (Gly) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATG (Met) | GTA (Val) |
HCM50/MK357732 | GGA (Gly) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATG (Met) | GTA (Val) |
DN8/MK357721 | GGA (Gly) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATG (Met) | GTA (Val) |
DN18/MK357722 | GGA (Gly) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATG (Met) | GTA (Val) |
DN26/MK357723 | GGA (Gly) | TTG (Leu) | TAT (Tyr) | GGT (Gly) | ACA (Thr) | AAC (Asn) | ATT (Ile) | GCT (Ala) | CCA (Pro) | CGA (Arg) | GAA (Glu) | ACA (Thr) | ATG (Met) | GTA (Val) |
Discussion
Study period | Region of Vietnam | Genotype of CPV-2 | References | ||
---|---|---|---|---|---|
2a | 2b | 2c | |||
2003–2004 | Northern and Southern | 0 | 8 | 1 | Nakamura et al., 2004 [16] |
2016–2018 | Northern, Central and Southern | 6 | 0 | 251 | This study |