التفاصيل البيبلوغرافية
| العنوان: |
RT-nested PCR detection of Mourilyan virus in Australian Penaeus monodon and its tissue distribution in healthy and moribund prawns. |
| المؤلفون: |
Cowley JA; CSIRO Livestock Industries, Queensland Bioscience Precinct, 306 Carmody Road, St. Lucia 4067, Australia. jeff.cowley@csiro.au, McCulloch RJ, Rajendran KV, Cadogan LC, Spann KM, Walker PJ |
| المصدر: |
Diseases of aquatic organisms [Dis Aquat Organ] 2005 Sep 05; Vol. 66 (2), pp. 91-104. |
| نوع المنشور: |
Comparative Study; Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: |
English |
| بيانات الدورية: |
Publisher: Inter-Research Country of Publication: Germany NLM ID: 8807037 Publication Model: Print Cited Medium: Print ISSN: 0177-5103 (Print) Linking ISSN: 01775103 NLM ISO Abbreviation: Dis Aquat Organ Subsets: MEDLINE |
| أسماء مطبوعة: |
Original Publication: Oldendorf/Luhe, Federal Republic of Germany : Inter-Research, c1985- |
| مواضيع طبية MeSH: |
Bunyaviridae/*genetics , Penaeidae/*virology , Reverse Transcriptase Polymerase Chain Reaction/*methods, Animals ; Australia ; DNA Primers ; Gills/ultrastructure ; Gills/virology ; Hemocytes/virology ; In Situ Hybridization ; Microscopy, Electron, Transmission ; Sensitivity and Specificity |
| مستخلص: |
Mourilyan virus (MoV) is a newly identified virus of Penaeus monodon prawns that is genetically related to the Uukuniemi virus and other phleboviruses of the Bunyaviridae. This paper describes an RT-nested PCR test that can reliably detect between 2 and 6 copies of a synthetic MoV RNA. Total RNA isolated from the lymphoid organ, gills and haemocytes of P. monodon with moderate infections gave comparable amplicon yields in the RT-PCR step of the test. However, in prawns with extremely low-level infections, haemocytes and gill tissue proved slightly more reliable in detecting MoV RNA following nested PCR. The distribution of MoV in tissues of healthy and moribund P. monodon was examined by in situ hybridisation (ISH) using a digoxigenin-labelled DNA probe to a approximately 0.8 kb M RNA segment cDNA insert in clone pMoV4.1. The DNA probe targeted a region in the MoV M RNA segment containing a coding sequence with homology to the C-terminus of the G2 glycoprotein of phleboviruses. In healthy prawns harbouring an unapparent MoV infection, ISH signal primarily occurred in the lymphoid organ, where it was more prominent in hypertrophied cells of 'spheroids' than within cells of normal tubules. ISH signal was also sometimes detected in cells of cuticular epithelium, segmental nerve ganglion and the antennal and tegmental glands. MoV was distributed widely throughout these and other cephalothoracic tissues of mesodermal and ectodermal origin in moribund P. monodon following experimental infection or collected from farm pond edges during disease episodes. Transmission electron microscopy of gill of moribund, captive-reared P. monodon identified spherical (approximately 85 nm diameter) to ovoid MoV particles (approximately 85 x 100 nm) in and around highly necrotic cells in which the nucleus and other organelles had disintegrated. MoV virions co-existed with rod-shaped virions of gill-associated virus and were often seen clustered within cytoplasmic vacuoles or associated with the outer rim of concentric ring-shaped structures comprised of endoplasmic membranes likely to represent degenerated Golgi. |
| المشرفين على المادة: |
0 (DNA Primers) |
| تواريخ الأحداث: |
Date Created: 20051020 Date Completed: 20060328 Latest Revision: 20061115 |
| رمز التحديث: |
20231215 |
| DOI: |
10.3354/dao066091 |
| PMID: |
16231634 |
| قاعدة البيانات: |
MEDLINE |
