Nawagitgul P, Morozov I, Sirinarumitr T, Sorden S D, Paul P S
Department of Veterinary Microbiology and Preventive Medicine, Veterinary Medical Research Institute (VMRI), 1802 Elwood Drive, Ames, IA 50011-1240, USA.
Vet Microbiol. 2000 Jul 3;75(1):83-9. doi: 10.1016/s0378-1135(00)00199-1.
Porcine circovirus type 1 (PCV1), a PK-15 cell line contaminant, and porcine circovirus type 2 (PCV2), associated with post-weaning multisystemic wasting syndrome (PMWS), are genetically and antigenically related. Several techniques have been developed to detect PCV, including in situ hybridization (ISH). Previously reported probes used for ISH may hybridize with both PCV1 and PCV2 nucleic acids. We attempted to produce probes for ISH that can detect and differentiate PCV2 from PCV1 in PCV-infected cells. Riboprobes were synthesized from the sense and antisense strands of both open reading frames 1 and 2 (ORF1 and ORF2) of PCV2. At 42 and 58 degrees C, the ORF1 antisense probe hybridized with nucleic acid from both PCV1- and PCV2-infected cells. At 58 degrees C, the ORF2 antisense probe hybridized with PCV2 nucleic acid but not with PCV1 nucleic acid. The ORF1 and ORF2 sense probes bound only with PCV2 nucleic acid. Both antisense strand probes produced stronger signals than the sense strand probes. The results showed that the PCV2 ORF1 antisense probe is the most likely probe to detect both PCV types while the ORF2 antisense probe is capable of discriminating between PCV1 and PCV2.
1型猪圆环病毒（PCV1）是PK - 15细胞系的污染物，与断奶后多系统衰竭综合征（PMWS）相关的2型猪圆环病毒（PCV2）在遗传和抗原方面存在关联。已经开发了多种检测PCV的技术，包括原位杂交（ISH）。先前报道的用于ISH的探针可能会与PCV1和PCV2核酸都发生杂交。我们试图制备用于ISH的探针，以便在PCV感染的细胞中检测PCV2并将其与PCV1区分开来。从PCV2的开放阅读框1和2（ORF1和ORF2）的有义链和反义链合成了核糖探针。在42℃和58℃时，ORF1反义探针与PCV1和PCV2感染细胞的核酸都发生杂交。在58℃时，ORF2反义探针与PCV2核酸杂交，但不与PCV1核酸杂交。ORF1和ORF2有义探针仅与PCV2核酸结合。两条反义链探针产生的信号都比有义链探针更强。结果表明，PCV2 ORF1反义探针最有可能同时检测两种PCV类型，而ORF2反义探针能够区分PCV1和PCV2。
