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Creation of Standard DNA for Detection and Quantification of White Spot Syndrome Virus in Shrimps by Real-time PCR (qPCR)

Authors :
Nguyen Thi Hong Loan
Phan Tuan Nghia
Tran Viet Cuong
Source :
VNU Journal of Science: Natural Sciences and Technology.
Publication Year :
2022
Publisher :
Vietnam National University Journal of Science, 2022.

Abstract

White spot syndrome virus (WSSV) is a double stranded DNA virus that causes WSS diseases for many crustaceans including the Penaeidae family shrimp. In this study, standard DNA for detection and quantification of WSSV by real-time polymerase chain reaction (qPCR) method was created. A specific gene fragment of 126 bp from WSSV genome was successfully amplified by PCR and cloned into pGEM-T vector. qPCR calibration curves using the created recombinant pGEM vector harboring 126 bp gene fragment as the standard DNA at concentrations of 3x102 to 3x108 copies/mL showed to have good linear regression with an efficiency of 99.1%, correlation coefficient R2 of 0.998 and the slope of -3.344. The recombinant vector was also used as a positive standard for detection and quantification of WSSV in a number of WSSV-infected shrimp samples and the values ranging from 3.69x103 copies/mL to 1.25x108 copies/mL WSSV were found in the collected samples.

Subjects

Subjects :
General Medicine

Details

ISSN :
25881140 and 26159317
Database :
OpenAIRE
Journal :
VNU Journal of Science: Natural Sciences and Technology
Accession number :
edsair.doi...........c4cd793eb6ab9f067a0e59bbd38f6260