Establishment of in situ hybridization technique for detection of porcine circovirus type 3 antigen in processed lung tissue and lymphoid organs

Lee, Kah Chun (2020) Establishment of in situ hybridization technique for detection of porcine circovirus type 3 antigen in processed lung tissue and lymphoid organs. [Project Paper] (Submitted)

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Abstract

Porcine circovirus type 3 (PCV3) is a newly emerging virus that has been recently reported in different countries. PCV3 has been associated with different clinical manifestations which include porcine dermatitis and nephropathy syndrome (PDNS), porcine respiratory disease complex (PDRC), reproductive failure and multisystemic inflammation. However, previous study on detection methods of PCV3 were limited in Malaysia. Hence, the current study aimed to detect the presence of PCV3 antigen in lung tissue and lymphoid organs via in situ hybridization (ISH). Using convenient sampling method, 10 archived homogenized lung samples and lymphoid organs were selected respectively for this study. The samples were previously obtained from clinically ill weaned pigs with the age ranges from 4-8 weeks old, and presented with clinical signs of weakness, sick, runt, moribund and having the history of respiratory distress. The samples were then subjected to conventional PCR assay using specific primer targeting the cap gene (ORF2), followed by hematoxylin and eosin staining (H&E) and subsequently ISH. Based on the PCR assay, 30% (3/10) of lungs were positive for PCV3 while 40% (4/10) of mesenteric lymph node and 40% (4/10) of inguinal lymph node were positive for PCV3 respectively. Upon H&E, the common clinical findings of PCV3 positive lungs are vessels congestions and infiltration of mononuclear cells at alveolar septa and alveolar cavity, while the common findings of PCV3 positive lymphoid organs are vessels congestion and depletion of lymphoid cells. Furthermore, brown PCV3 antigens are detected in 100% (3/3) of lung tissues via ISH while the positive detection rate of mesenteric lymph node and inguinal lymph node are 100% (4/4) and 75% (3/4) respectively. In conclusion, this study had successfully detected PCV3 antigen in processed lung tissue and lymphoid organs via ISH. However, the sample size used for this study is too small and larger sample size should be included in future studies.

Item Type:	Project Paper
Faculty:	Faculty of Veterinary Medicine
Depositing User:	Ms. Nordeena Abdul Aziz
Date Deposited:	18 Oct 2023 08:50
Last Modified:	18 Oct 2023 08:50
URI:	http://psaspb.upm.edu.my/id/eprint/1457

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