Tags

Type your tag names separated by a space and hit enter

Detection and localization of naturally transmitted avian leukosis subgroup J virus in egg-type chickens by in situ PCR hybridization.
J Vet Med A Physiol Pathol Clin Med. 2007 Dec; 54(10):553-8.JV

Abstract

Avian leukosis virus (ALV) subgroup J (ALV-J) is an exogenous ALV and causes myeloid leukosis in meat-type chickens. We have previously reported the isolation and identification of ALV-J in commercial layer flocks from 12 farms in northern China. In this report, we further characterized this virus by in situ polymerase chain reaction (PCR) hybridization in various affected organs of chickens from six of the 12 farms. A routine method for hybridization of nucleic acid uses radioactive probe, such as a P32-labelled probe. We found that the non-radioactive digoxigenin (DIG) probe is sensitive enough to detect the nucleic acid of virus in chicken tissues. We used a pair of published primers (H5/H7) specific to the gp85 envelope gene and 3' region of pol gene of prototype ALV-J strain HPRS-103. The total RNA extracted from tumour, bone marrow, oviduct, liver and spleen of the diseased chickens from six commercial flocks, and cDNA was successfully amplified. Using the primers and cDNA, we obtained an ALV-J-specific cDNA probe of 545 bp in length by PCR. In situ PCR with H5/H7 primers was carried out in the paraffin sections from tissues of the diseased chickens, followed by in situ hybridization using the DIG-labelled cDNA probe. Positive hybridization signals were detected in the cytoplasm of paraffin sections of tumours and other organ tissues. The intensity of the signals was documented using an image analysis system measuring integral optical density (IOD). The IOD values for tissue sections treated by in situ PCR hybridization are significantly higher than that by in situ hybridization alone (P < 0.01). These data taken together suggest that in situ PCR hybridization is a more sensitive technique for detection of ALV-J in tissue sections.

Authors+Show Affiliations

College of Veterinary Medicine, China Agricultural University, Beijing 100094, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18045338

Citation

Li, N, et al. "Detection and Localization of Naturally Transmitted Avian Leukosis Subgroup J Virus in Egg-type Chickens By in Situ PCR Hybridization." Journal of Veterinary Medicine. A, Physiology, Pathology, Clinical Medicine, vol. 54, no. 10, 2007, pp. 553-8.
Li N, Xu B, Dong W, et al. Detection and localization of naturally transmitted avian leukosis subgroup J virus in egg-type chickens by in situ PCR hybridization. J Vet Med A Physiol Pathol Clin Med. 2007;54(10):553-8.
Li, N., Xu, B., Dong, W., Qiao, S., Lee, L. F., Zhang, H. M., Li, M., & Du, N. (2007). Detection and localization of naturally transmitted avian leukosis subgroup J virus in egg-type chickens by in situ PCR hybridization. Journal of Veterinary Medicine. A, Physiology, Pathology, Clinical Medicine, 54(10), 553-8.
Li N, et al. Detection and Localization of Naturally Transmitted Avian Leukosis Subgroup J Virus in Egg-type Chickens By in Situ PCR Hybridization. J Vet Med A Physiol Pathol Clin Med. 2007;54(10):553-8. PubMed PMID: 18045338.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Detection and localization of naturally transmitted avian leukosis subgroup J virus in egg-type chickens by in situ PCR hybridization. AU - Li,N, AU - Xu,B, AU - Dong,W, AU - Qiao,S, AU - Lee,L F, AU - Zhang,H M, AU - Li,M, AU - Du,N, PY - 2007/11/30/pubmed PY - 2008/2/20/medline PY - 2007/11/30/entrez SP - 553 EP - 8 JF - Journal of veterinary medicine. A, Physiology, pathology, clinical medicine JO - J Vet Med A Physiol Pathol Clin Med VL - 54 IS - 10 N2 - Avian leukosis virus (ALV) subgroup J (ALV-J) is an exogenous ALV and causes myeloid leukosis in meat-type chickens. We have previously reported the isolation and identification of ALV-J in commercial layer flocks from 12 farms in northern China. In this report, we further characterized this virus by in situ polymerase chain reaction (PCR) hybridization in various affected organs of chickens from six of the 12 farms. A routine method for hybridization of nucleic acid uses radioactive probe, such as a P32-labelled probe. We found that the non-radioactive digoxigenin (DIG) probe is sensitive enough to detect the nucleic acid of virus in chicken tissues. We used a pair of published primers (H5/H7) specific to the gp85 envelope gene and 3' region of pol gene of prototype ALV-J strain HPRS-103. The total RNA extracted from tumour, bone marrow, oviduct, liver and spleen of the diseased chickens from six commercial flocks, and cDNA was successfully amplified. Using the primers and cDNA, we obtained an ALV-J-specific cDNA probe of 545 bp in length by PCR. In situ PCR with H5/H7 primers was carried out in the paraffin sections from tissues of the diseased chickens, followed by in situ hybridization using the DIG-labelled cDNA probe. Positive hybridization signals were detected in the cytoplasm of paraffin sections of tumours and other organ tissues. The intensity of the signals was documented using an image analysis system measuring integral optical density (IOD). The IOD values for tissue sections treated by in situ PCR hybridization are significantly higher than that by in situ hybridization alone (P < 0.01). These data taken together suggest that in situ PCR hybridization is a more sensitive technique for detection of ALV-J in tissue sections. SN - 0931-184X UR - https://www.unboundmedicine.com/medline/citation/18045338/Detection_and_localization_of_naturally_transmitted_avian_leukosis_subgroup_J_virus_in_egg_type_chickens_by_in_situ_PCR_hybridization_ L2 - https://doi.org/10.1111/j.1439-0442.2007.01008.x DB - PRIME DP - Unbound Medicine ER -