Detection of infectious hypodermal and hematopoietic necrosis virus (IHHNV) in Litopenaeus vannamei by ramification amplification assay

Diseases of Aquatic Organisms

Hung-Wen Chien¹  Man-Shu Chen¹  Fu-Chun Lee¹  Pei-Yu Lee¹  Ping-Hua Teng¹  Bor-Rung Ou¹  Ping-Feng Sung¹  Chen Su¹ 

关键词: IHHNV;    Litopenaeus vannamei;    Ramification amplification;    Isothermal amplification;   

DOI  :  10.3354/dao073103

学科分类：生物科学（综合）

来源: Inter-Research

【 摘 要 】

ABSTRACT: Infectious hypodermal and hematopoietic necrosis virus (IHHNV) is a single-stranded DNA virus that causes developmental and growth abnormalities in Pacific white shrimp Litopenaeus vannamei (also known as Penaeus vannamei). Nucleic acid based methods such as in situ hybridization (ISH) and PCR have been commonly used for IHHNV detection. Ramification amplification (RAM), an isothermal nucleic acid amplification approach, was used in this study to detect IHHNV in L. vannamei. RAM offers many advantages over PCR, including simple procedures and short detection time, and is labor-saving and cost-effective. RAM exponentially amplifies a circular oligonucleotide amplicon (C probe) after a target-specific ligation step through sequential primer extension and strand displacement processes. The conditions of an IHHNV RAM assay were optimized using artificial templates and targets prior to application. Using DNA of IHHNV-infected L. vannamei as targets, results revealed that RAM amplified target DNA with similar sensitivity as PCR. RAM offers competitive levels of speed, simplicity and sensitivity among various pathogen diagnostic methods.

【 授权许可】

Unknown