Sensitive and rapid detection of infectious hypodermal and hematopoietic necrosis virus (IHHNV) in shrimps by loop-mediated isothermal amplification

Sun, Zhaofeng; Hu, Chaoqun; Ren, Chen; Shen, Qi

doi:10.1016/j.jviromet.2005.07.011

Cited by 64 publications

(28 citation statements)

References 13 publications

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“…LAMP has been successfully applied for rapid and real-time detection of both DNA and RNA viruses. Recently, LAMP has been used for detection of shrimp viruses such as white spot syndrome virus (WSSV) infecting M. japonicus [15], infection of infectious hypodermal and hematopoietic necrosis virus (IHHNV) in shrimps [16], and yellow head virus (YHV) in shrimps [17]. In addition, LAMP has been also reported for the detection of virus in fish such as seabream iridovirus [18], koi herpesvirus (KHV) [19], infectious hematopoietic necrosis virus (IHNV) in rainbow trout [20], viral hemorrhagic septicemia (VHS) of salmonid fish [21], and spring viremia of carp (SVC) [22].…”

Section: Discussionmentioning

confidence: 99%

“…The present Q-LAMP assay can be used as an alternative to conventional PCR and routine LAMP assay [36]. Compared to other quantification methods, the Loopamp turbidimeter is comparatively less expensive than Q-PCR, which requires fluorogenic primers and probes [16]. The most beneficial property of the V. nigripulchritudo Q-LAMP assay is its rapidity; detection can be completed within a short duration, whereas conventional PCR methods require 2-3 h [37].…”

Section: Discussionmentioning

confidence: 99%

“…16 20 24 28 32 36 40 44 48 52 56 60 64 68 72 76 80 Reaction time (min) Optimization of the reaction temperature for V. nigripulchritudo Q-LAMP performed at 60, 63, and 65°C.…”

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confidence: 99%

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Quantitative loop-mediated isothermal amplification method for the detection of Vibrio nigripulchritudo in shrimp

et al. 2010

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Vibrio nigripulchritudo is considered one of the major pathogens threatening shrimp aquaculture. In this study, we developed a novel and highly specific quantitative loop-mediated isothermal amplification (Q-LAMP) assay. A set of four specific primers were designed targeting the V. nigripulchritudo intergenic spacer region. The reaction time and temperature were optimized for 60 min at 63°C. Quantitative analysis was then performed by measuring the turbidity of the reaction solution using a real-time turbidimeter, allowing for quantification of the initial DNA concentration with a sensitivity of 10 2 copy numbers equivalent to 2.3 colony forming units/ml or 0.3 fg/ll. The LAMP assay was able to specifically detect two representative strains of V. nigripulchritudo, whereas other Vibrio and non-Vibrio species were not amplified. A standard curve was generated for V. nigripulchritudo by plotting the threshold time (T t ) versus the log of bacterial number. A high correlation coefficient (R 2 = 0.9749) was observed for the Q-LAMP reaction. In conclusion, Q-LAMP assay is a sensitive, rapid, and simple tool that can be used for the detection and quantification of V. nigripulchritudo in shrimp, thereby facilitating surveillance of vibriosis infection.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Quantitative loop-mediated isothermal amplification method for the detection of Vibrio nigripulchritudo in shrimp

et al. 2010

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“…In this method, in addition to the four sets of primers normally used, one extra set of primer was required to accelerate the LAMP reaction. The technique developed in this study was faster than the usual LAMP reaction described in earlier study [77]. A real-time LAMP assay was developed for the detection of IHHNV infection in penaeid shrimps by employing four primers …”

Section: Loop-mediated Isothermal Amplificationmentioning

confidence: 99%

“…In 2006, a diagnostic LAMP based method was developed for IHHNV detection and the four sets of primers were designed targeting the IHHNV genome sequence (GenBank Accession No. AF218266) [77]. The target DNA is amplified and visualized using gel electrophoresis within 60 min under isothermal condition at 64°C.…”

Section: Loop-mediated Isothermal Amplificationmentioning

confidence: 99%

Genomics, Molecular Epidemiology and Diagnostics of Infectious hypodermal and hematopoietic necrosis virus

et al. 2012

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Infectious hypodermal and hematopoietic necrosis virus (IHHNV) is one of the major viral pathogens of penaeid shrimps worldwide, which has resulted in severe mortalities of up to 90 % in cultured Penaeus (Litopenaeus) stylirostris from Hawaii and hence designated Penaeus stylirostris densovirus (PstDNV). IHHNV is distributed in shrimp culture facilities worldwide. It causes large economic loss to the shrimp farming industry. Our knowledge about the natural reservoirs of IHHNV is still scarce. Recent studies suggest that there is sufficient sequence variation among the isolates from different locations in Asia, suggesting multiple geographical strains of the virus. Four complete genomes and several partial sequences of the virus are available in the GenBank. Complete genome information would be useful for assessing the specificity of diagnostics for viruses from different geographical areas. Comparisons of complete genome sequences will help us gain insights into point mutations that can affect virulence of the virus. In addition, because of unavailability of shrimp cell lines for culturing IHHNV in vitro, quantification of virus is difficult. The recent progress in research regarding clinical signs, geographical distribution, complete genome sequence and genetic variation, transmission has made it possible to obtain information on IHHNV. A comprehensive understanding of IHHNV infection process, pathogenesis, structural proteins and replication is essential for developing prevention measures. To date, no effective prophylactic measure for IHHNV infection is available for shrimp to reduce its impact. This review provides an overview of key issues regarding IHHNV infection and disease in commercially important shrimp species.

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Complexities of Diagnostics of Viruses Affecting Farmed Aquatic Species

Weidmann

2017

Diagnosis and Control of Diseases of Fish and Shellfish

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Sensitive and rapid detection of infectious hypodermal and hematopoietic necrosis virus (IHHNV) in shrimps by loop-mediated isothermal amplification

Cited by 64 publications

References 13 publications

Quantitative loop-mediated isothermal amplification method for the detection of Vibrio nigripulchritudo in shrimp

Quantitative loop-mediated isothermal amplification method for the detection of Vibrio nigripulchritudo in shrimp

Genomics, Molecular Epidemiology and Diagnostics of Infectious hypodermal and hematopoietic necrosis virus

Complexities of Diagnostics of Viruses Affecting Farmed Aquatic Species

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