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:: Volume 27, Issue 6 (3-2019) ::
isfj 2019, 27(6): 115-125 Back to browse issues page
The introduce of a new modified Nested RT-PCR molecular technique for rapid diagnosis of Viral Nervous Necrosis (VNN)
F. Hassantabar1 , F. Firouzbakhsh1 , M.J. Zorriehzahra2 , K. D. Thompson3
1- Sari Agricultural Sciences and Natural Resources University
2- Aquatic Animal Health and Diseases Dept., Iranian Fisheries Science Research Institute (IFSRI), Agricultural Research Education and Extension Organization (AREEO), Tehran, Iran
3- Moredun Research Institute
Abstract:   (5320 Views)
Viral Nervous Necrosis (VNN) is considered as one of the most important threats for mariculture and aquaculture in the world and up to now, it has been affecting more than 70 fish species. This study was aimed to detect and identify fish betanodavirus, the causative agent of VNN, using nested RT-PCR from brain tissue of Caspian Sea Mullet. Forty samples of subclinical golden grey mullet fish (range: 50-250 gr) were collected from the coastal areas of Caspian Sea in 2016. The target organ (brain) was excised and the reverse transcription (cDNA synthesis) was carried out followed by the RNA extraction. In the present study, detecting of the virus was performed with Nested RT-PCR using two pairs of primers F2/R3 and NF2/NR3 in two steps. After optimization of PCR, all of the forty specimens were negative by the first-step PCR (RT-PCR) while 20 out of 40 grey mullet were positive using Nested RT-PCR (second-step PCR) and a strong band of the expected size (300 bp) was clearly observed. Therefore, it seems that Nested RT-PCR is a method of choice for rapid detection of the low level of infection and in asymptomatic carrier fish. In addition, the result of sequencing revealed that the virus belonged to the RGNNV genotype.

 
 

Keywords: Viral Nervous Necrosis, Golden grey mullet, Nested RT-PCR, Sequencing
Full-Text [PDF 814 kb]   (3611 Downloads)    
Type of Study: Research | Subject: بهداشت بيماريها
Received: 2018/04/25 | Published: 2019/03/15
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Hassantabar F, Firouzbakhsh F, Zorriehzahra M, D. Thompson K. The introduce of a new modified Nested RT-PCR molecular technique for rapid diagnosis of Viral Nervous Necrosis (VNN). isfj 2019; 27 (6) :115-125
URL: http://isfj.ir/article-1-2044-en.html


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Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Volume 27, Issue 6 (3-2019) Back to browse issues page

با کسب مجوز از دفتر کمیسیون بررسی نشریات علمی وزارت علوم، تحقیات و فنآوری مجله علمی شیلات بصورت آنلاین می باشد و تعداد محدودی هم به چاپ می رساند. شماره شاپای جدید آن ISSN:2322-5998 است

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