Development and evaluation of an indirect in situ polymerase chain reaction for the detection of porcine circovirus type 2 in formalin-fixed and paraffin-embedded tissue specimens
| Autor: | Chen-Hsuan Liu, Chian-Ren Jeng, Victor Fei Pang, Mi Yuan Chia, Yi Chieh Tsai, Chun-Ming Lin, Chih-Cheng Chang, Shih Hsuan Hsiao |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Circovirus
Pathology medicine.medical_specialty Swine animal diseases In situ hybridization Biology Polymerase Chain Reaction Sensitivity and Specificity Microbiology law.invention Porcine Postweaning Multisystemic Wasting Syndrome law Formaldehyde medicine Animals Polymerase chain reaction Paraffin Embedding General Veterinary virus diseases General Medicine biology.organism_classification Staining Porcine circovirus Immunohistochemistry Lymph Nodes Tissue Preservation Circoviridae Nested polymerase chain reaction |
| Zdroj: | Veterinary Microbiology. 138:225-234 |
| ISSN: | 0378-1135 |
| DOI: | 10.1016/j.vetmic.2009.04.002 |
| Popis: | Taking advantage of the high sensitivity of polymerase chain reaction (PCR) and the cell-localizing ability of in situ hybridization (ISH), an indirect in situ PCR (ISPCR) method was developed for detecting the distribution of porcine circovirus type 2 (PCV2) in formalin-fixed and paraffin-embedded inguinal lymph nodes obtained from clinically healthy PCV2-carrier pigs and postweaning multisystemic wasting syndrome (PMWS)-affected pigs. Comparisons of the relative sensitivity of indirect ISPCR with other routinely used diagnostic methods for PCV2 indicated that nested PCR was the most sensitive method followed by indirect ISPCR, conventional PCR, ISH, and immunohistochemical (IHC) staining. Although indirect ISPCR, ISH, and IHC staining all revealed a similar signal distribution pattern of PCV2, using indirect ISPCR allowed specific amplification and detection of previously uneasily detected PCV2 signal than by routine ISH or IHC staining, particularly in those cells within the germinal center in clinically healthy PCV2-carrier pigs. Furthermore, six different PCV2 signal expression patterns in conjunction with the correlated lymphoid lesion stages were classified to describe the tissue morphological changes and viral infection. The result indicates that indirect ISPCR is a more effective, cell-based diagnostic tool with good specificity to detect limited PCV2 infection in formalin-fixed and paraffin-embedded tissue specimens and it would be a useful tool for further exploring the pathogenesis of PCV2 infection. |
| Databáze: | OpenAIRE |
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