Design and Evaluation of a PCR Method for Detecting White Spot Syndrome Virus in Shrimp

Background: White spot syndrome virus (wssv) is the causing agent for white spot disease in shrimp and many crustaceans. This disease is highly contagious and can cause death within 3–10 days under normal culture conditions. Therefore, early diagnosis of the virus is a necessity. Materials and Metho...

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Autores principales:	Hossein Moein, Farshid Kafilzadeh, Mohammad Reza Miri, Mohammad Khalilpazir
Formato:	article
Lenguaje:	EN FA
Publicado:	Bushehr University of Medical Sciences 2021
Materias:	white spot syndrome virus pcr diagnosis Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/5e30c159dad44625ac3eff60b139992a
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spelling	oai:doaj.org-article:5e30c159dad44625ac3eff60b139992a2021-12-04T05:40:39ZDesign and Evaluation of a PCR Method for Detecting White Spot Syndrome Virus in Shrimp1735-43741735-6954https://doaj.org/article/5e30c159dad44625ac3eff60b139992a2021-11-01T00:00:00Zhttp://ismj.bpums.ac.ir/article-1-1516-en.htmlhttps://doaj.org/toc/1735-4374https://doaj.org/toc/1735-6954Background: White spot syndrome virus (wssv) is the causing agent for white spot disease in shrimp and many crustaceans. This disease is highly contagious and can cause death within 3–10 days under normal culture conditions. Therefore, early diagnosis of the virus is a necessity. Materials and Methods: Primers were designed for three regions of the virus genome and one region of the shrimp genome so that they could function together in a reaction. DNA was extracted from 40 samples of shrimp suspected of white spot disease. Primers were then optimized individually to detect white spot virus and after selecting the most suitable one, the virus was detected simultaneously by two pairs of primers. Shrimp genome replication primers were also used as internal control. Results: Among the designed primers, three pairs of primers were selected that amplified one fragment of the shrimp genome and two fragments of the virus genome. Of the 40 samples examined, 28 samples were positive (infected with the virus) and 12 samples were negative, which completely matched the results obtained with the reference kit. Conclusion: To detect white spot virus, examination of two regions of the virus genome is sufficient and reduces the possibility of false negatives. It is also effective to use the shrimp genome to control DNA extraction and PCR steps. The single-step method (PCR) is preferable to the two-step method (Nested PCR) due to its reduced probability of contamination and ease of use.Hossein MoeinFarshid KafilzadehMohammad Reza MiriMohammad KhalilpazirBushehr University of Medical Sciencesarticlewhite spot syndrome viruspcrdiagnosisMedicine (General)R5-920ENFAIranian South Medical Journal , Vol 24, Iss 5, Pp 469-480 (2021)
institution	DOAJ
collection	DOAJ
language	EN FA
topic	white spot syndrome virus pcr diagnosis Medicine (General) R5-920
spellingShingle	white spot syndrome virus pcr diagnosis Medicine (General) R5-920 Hossein Moein Farshid Kafilzadeh Mohammad Reza Miri Mohammad Khalilpazir Design and Evaluation of a PCR Method for Detecting White Spot Syndrome Virus in Shrimp
description	Background: White spot syndrome virus (wssv) is the causing agent for white spot disease in shrimp and many crustaceans. This disease is highly contagious and can cause death within 3–10 days under normal culture conditions. Therefore, early diagnosis of the virus is a necessity. Materials and Methods: Primers were designed for three regions of the virus genome and one region of the shrimp genome so that they could function together in a reaction. DNA was extracted from 40 samples of shrimp suspected of white spot disease. Primers were then optimized individually to detect white spot virus and after selecting the most suitable one, the virus was detected simultaneously by two pairs of primers. Shrimp genome replication primers were also used as internal control. Results: Among the designed primers, three pairs of primers were selected that amplified one fragment of the shrimp genome and two fragments of the virus genome. Of the 40 samples examined, 28 samples were positive (infected with the virus) and 12 samples were negative, which completely matched the results obtained with the reference kit. Conclusion: To detect white spot virus, examination of two regions of the virus genome is sufficient and reduces the possibility of false negatives. It is also effective to use the shrimp genome to control DNA extraction and PCR steps. The single-step method (PCR) is preferable to the two-step method (Nested PCR) due to its reduced probability of contamination and ease of use.
format	article
author	Hossein Moein Farshid Kafilzadeh Mohammad Reza Miri Mohammad Khalilpazir
author_facet	Hossein Moein Farshid Kafilzadeh Mohammad Reza Miri Mohammad Khalilpazir
author_sort	Hossein Moein
title	Design and Evaluation of a PCR Method for Detecting White Spot Syndrome Virus in Shrimp
title_short	Design and Evaluation of a PCR Method for Detecting White Spot Syndrome Virus in Shrimp
title_full	Design and Evaluation of a PCR Method for Detecting White Spot Syndrome Virus in Shrimp
title_fullStr	Design and Evaluation of a PCR Method for Detecting White Spot Syndrome Virus in Shrimp
title_full_unstemmed	Design and Evaluation of a PCR Method for Detecting White Spot Syndrome Virus in Shrimp
title_sort	design and evaluation of a pcr method for detecting white spot syndrome virus in shrimp
publisher	Bushehr University of Medical Sciences
publishDate	2021
url	https://doaj.org/article/5e30c159dad44625ac3eff60b139992a
work_keys_str_mv	AT hosseinmoein designandevaluationofapcrmethodfordetectingwhitespotsyndromevirusinshrimp AT farshidkafilzadeh designandevaluationofapcrmethodfordetectingwhitespotsyndromevirusinshrimp AT mohammadrezamiri designandevaluationofapcrmethodfordetectingwhitespotsyndromevirusinshrimp AT mohammadkhalilpazir designandevaluationofapcrmethodfordetectingwhitespotsyndromevirusinshrimp
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Design and Evaluation of a PCR Method for Detecting White Spot Syndrome Virus in Shrimp

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