Detection of yellow head virus in shrimp by loop-mediated isothermal amplification (LAMP)

Mekata, Tohru; Kono, Tomoya; Savan, Ram; Sakai, Masahiro; Kasornchandra, Jiraporn; Yoshida, Terutoyo; Itami, Toshiaki

doi:10.1016/j.jviromet.2006.02.012

Cited by 59 publications

(30 citation statements)

References 15 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%

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%

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

et al. 2010

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“…LAMP is a novel method of DNA amplification that has already been applied to detection of several pathogens in aquaculture (Savan et al 2005). These include IHNV (Gunimaladevi et al 2005, McCarthy et al 2006, infectious salmon anemia virus (ISAV) (McCarthy et al 2006), VHSV (Soliman & ElMatbouli 2006), spring viraemia of carp virus (SVCV) (Liu et al 2008), IPNV (Soliman et al 2009), turbot reddish body iridovirus (TRBIV) (Zhang et al 2009), white spot syndrome virus (WSSV) , koi herpesvirus (KHV) (Gunimaladevi et al 2004), yellow head virus (YHV) (Mekata et al 2006) and Taura syndrome virus (TSV) (Kiatpathomchai et al 2007). According to these studies, the sensitivity of the LAMP assay is higher than the conventional PCR assay.…”

Section: Discussionmentioning

confidence: 99%

Reverse transcriptase loop-mediated isothermal amplification assay for infectious hematopoietic necrosis virus in Oncorhynchus keta

Suebsing¹,

Jeon²,

Oh³

et al. 2011

Dis. Aquat. Org.

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A reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay was developed for detecting infectious hematopoietic necrosis virus (IHNV) from chum salmon Oncorhynchus keta in South Korea with high specificity, sensitivity and rapidity. A set of 6 IHNV-specific primers was designed, based on the G-protein sequence of IHNV (PRT strain), recognizing 8 distinct sequences of the target RNA. The assay was optimized to detect IHNV at 63°C for 30 min. The limit of detection was 0.01 fg of RNA extracted from IHNV-infected CHSE-214 cells, compared with 1.0 fg for nested RT-PCR. The applicability of this RT-LAMP assay was further tested by comparison with nested RT-PCR using field samples. Of 473 samples tested, 191 samples (40.38%) were IHNVpositive by RT-LAMP, whereas 162 samples (34.25%) were IHNV-positive by nested RT-PCR. These results indicate that, because of its high sensitivity and rapidity, the RT-LAMP assay is useful for early diagnosis of IHN. KEY WORDS: RT-LAMP · IHNV · Chum salmon · Oncorhynchus keta Resale or republication not permitted without written consent of the publisherDis Aquat Org 94: [1][2][3][4][5][6][7][8] 2011 method. However, nested RT-PCR methods require a second amplification step that is prone to contamination. To solve these problems, the loop-mediated isothermal amplification (LAMP) assay was investigated as an alternative. LAMP was originally developed by Notomi et al. (2000) and can amplify very low numbers of target sequences to 10 9 copies under isothermal conditions within 1 h. This method depends on auto-cycling strand displacement DNA synthesis by the Bst DNA polymerase large fragment with high strand displacement activity, and a set of 2 specially designed inner primers and 2 outer primers. LAMP is highly specific because the target sequences are detected by 6 independent primers in the initial stage, followed by 4 independent primers in the later stages of the LAMP reaction. LAMP is also applicable for RNA detection by using a reverse transcriptase (RT) together with a DNA polymerase (Notomi et al. 2000). This technique can be carried out under isothermal conditions, which can be simply achieved with a water bath or a heating block. Expensive thermal cyclers used for PCR are not required (Notomi et al. 2000).In the present study, an RT-LAMP assay was developed for detecting IHNV from wild adult chum salmon Oncorhynchus keta and artificially hatched fry in Korea. A set of 6 LAMP-specific primers was designed, based on the G protein sequence of Korean IHNV (PRT strain, accession no. AY673684). The applicability of the RT-LAMP assay was evaluated by comparison with the nested RT-PCR assay using field samples. MATERIALS AND METHODSViruses and cell culture. IHNV (PRT strain) was amplified in chinook salmon embryo (CHSE)-214 cells. Infectious pancreatic necrosis virus (IPNV) (VR-299, Jasper, Sp and Ab strains) and viral hemorrhagic septicemia virus (VHSV) were maintained in rainbow trout gonad (RTG)-2 cells with minimum essential medium-10 (supplemented wit...

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“…), yellow head virus (Mekata et al . ), infectious hypodermal and hematopoietic necrosis virus (Sun et al . ), Taura syndrome virus (Kiatpathomchai et al .…”

Section: The Primer Set Used In This Studymentioning

confidence: 99%

Detection of acute hepatopancreatic necrosis disease strain of Vibrio parahaemolyticus using loop‐mediated isothermal amplification

Koiwai

Tinwongger

Nozaki

et al. 2015

Journal of Fish Diseases

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Detection of yellow head virus in shrimp by loop-mediated isothermal amplification (LAMP)

Cited by 59 publications

References 15 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

Reverse transcriptase loop-mediated isothermal amplification assay for infectious hematopoietic necrosis virus in Oncorhynchus keta

Detection of acute hepatopancreatic necrosis disease strain of Vibrio parahaemolyticus using loop‐mediated isothermal amplification

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