Development of in situ hybridization assay that differentiates between two genotypes of Porcine circovirus-2 in formalin-fixed, paraffin-embedded tissues.
The aim of the current study was to develop a nonradioactive in situ hybridization assay that can differentiate between genotypes 2a and 2b of Porcine circovirus-2 (PCV-2) in formalin-fixed, paraffin-embedded lymph node tissues from pigs with postweaning multisystemic wasting syndrome. Two different digoxigenin-labeled oligonucleotide probes were designed from the PCV-2 open reading frame 2 sequences. The PCV-2a-specific probe did not hybridize with formalin-fixed, paraffin-embedded lymph nodes from naturally PCV-2b-infected pigs and vice versa. Both PCV-2a-specific and PCV-2b-specific probes gave consistent negative signals in lymph nodes from naturally PCV-1-infected pigs. The in situ hybridization assay described in the present study represents a diagnostic tool that can differentiate between the 2 genotypes of PCV-2.
Kim
Duyeol
D
Department of Veterinary Pathology, College of Veterinary Medicine, Seoul National University, San 56-1, Shillim-dong, Gwanak-gu, Seoul, 151-742, Republic of Korea.
Ha
Yooncheol
Y
Oh
Yeonsu
Y
Han
Kiwon
K
Noh
Seung Ho
SH
Kim
Cheolwoo
C
Kim
Sung-Hoon
SH
Chae
Chanhee
C
D000818:Animals; D017925:Circovirus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D005557:Formaldehyde / Q000737:chemistry*; D005838:Genotype; D017403:In Situ Hybridization / Q000379:methods / Q000662:veterinary*; D016612:Paraffin Embedding / Q000662:veterinary*; D053570:Porcine Postweaning Multisystemic Wasting Syndrome / Q000821:virology*; D013552:Swine; D016707:Tissue Fixation / Q000662:veterinary*
D005557:Formaldehyde